Enoxaparin FDA 2010

('l
DEPARTMET OF HETH &. HUMA SERVICES

'"'''~'''~
Food and Drug Administration
Rockvile M D 20857
JUl 2 3 2010
Peter O. Safir
Scott L. Cunningham
Covington & Burling
1201 Pennsylvania Ave., N.W.
Washington, D.C. 20004-2401
Re: Docket No. FDA-2003-P-0273
Dear Mr. Safir and Mr. Cunningham:

This letter responds to your citizen petition submitted on behalf of A ventis Pharmaceuticals, Inc.
(Aventis) on February 19,2003 (Petition),! requesting that the Food and Drug Administration
(FDA or the Agency) withhold approval of any abbreviated new drug application (ANDA) for a
generic2 version of
Lovenox (enoxaparin sodium injection) (hereafter, "generic enoxaparin")
unless certain conditions are met. Specifically, you make the following requests:
That until enoxaparin has been fully characterized, we not approve any
ANDA citing Lovenox as the reference listed drug (RLD) unless the manufacturing process used to create the generic drug product is determined to be
equivalent to Aventis's manufacturing process for enoxaparin or the application is supported by proof of equivalent safety and effectiveness demonstrated
through clinical trials.
That we not approve any ANDA citing Lovenox as the RLD unless the generic product contains a 1,6 anhydro ring structure at the reducing ends of be3
tween 15 percent and 25 percent of its poly( oligo )saccharide chains.
We have carefully reviewed your petition and the supplements submitted by you on behalf of
Aventis and its successors in interest on February 12,2004 (Supplement No.1), September 26,
2005 (Supplement No.2), September 14,2006 (Supplement No.3), and June 29, 2007 (Supplement No.4), as well as the comments submitted by you (dated September 1,2004, October 13,
2004, March 17,2005, March 16,2006, August 25, 2006, March 2,2007, March 15,2007, and
i This citizen petition was originally assigned docket number 2003P-0064/CPI. The number was changed to FDA2003-P-0273 as a result of FDA's transition to its new docketing system (Regulations.gov) in January 2008.
2 The term "generic" refers to a drug product for which approval is sought under an ANDA submitted under section
the Federal Food, Drug, and Cosmetic Act (the Act).

5050) of
3 A monosaccharide is a simple sugar (carbohydrate) that cannot be decomposed by hydrolysis to obtain smaller
molecules of carbohydrate. A saccharide chain contains a series of monosaccharides linked by glycosidic bonds. A
many monosaccharides joined by glycosidic bonds. An
polysaccharide is a saccharide chain composed of
monosaccharides linked by glycosidic bonds.
oligosaccharide is a saccharide chain containing a small number of
The terms polysaccharide and oligosaccharide are sometimes used interchangeably.
Docket No. FDA-2003-P-0273
14, 2009). We have also reviewed the comments submitted by Amphastar Pharmaceuticals, Inc. (Amphastar) (dated May 13,2004, November 23,2004, July 18,2005, October 6,
2005, and October 7,2005), Hyman, Phelps & McNamara, P.C. (dated October 17,2003, and
August 4,2004), Teva Pharaceuticals USA Inc. (dated August 20, 2004), and other commentApril
ers. We have also extensively reviewed relevant scientific evidence and information.
For the reasons stated below, your petition is granted in par with respect to your request that
generic enoxaparin contain the 1,6 anydro ring structure at the reducing ends of between 15
percent and 25 percent of its poly( oligo )saccharide, but denied in all other respects.4
OVERVIEW
To obtain ANDA approval, an applicant must submit suffcient information to show that its
generic drug product is bioequivalent to and has the same active ingredient(s), route of adminiuse, and (with certain
stration, dosage form, strength, previously approved conditions of
the ANDA approval requirements
exceptions) labeling as the RLD. The underlying premise of
generic
drug
product
and
the
RLD
can
be
substituted
for each other with the full
is that the
expectation that they wil have the same clinical effect and safety profile.5
The primary issue raised by the petition and related comments is whether an ANDA applicant for
enoxaparin can demonstrate that its product has the same active ingredient (i.e., enoxaparin
sodium or "enoxaparin") as that of
your petition
the RLD (i.e., Aventis's Lovenox). The crux of
and related supplements is that we should not approve an ANDA forenoxaparin unless the
ANDA applicant: (1) completely characterizes enoxaparin by isolating, purifying, and sequencing each of its unique polysaccharide chains and determining their relative abundance, which you
state is curently impossible, (2) uses Aventis's or the equivalent manufacturing process, or (3)
conducts clinical trials to demonstrate the equivalent safety and effectiveness of the product.
We do not find it necessary for an ANDA applicant seeking approval of generic enoxaparin to
submit the information you request. Nonetheless, we recognize that the approval of ANDAs for
enoxaparin raises complicated scientific and regulatory issues, which we have carefully considbatch-
ered. Lovenox, a naturally sourced drug product, has some degree of
to-batch variabilty,
4 As discussed in section VI, you do not clarify in your petition and supplements what you mean by your request that
an ANDA applicant for enoxaparin use a manufacturing process equivalent to Aventis's manufacturing process. To
manufacture enoxaparin, an ANDA applicant generally (1) would depolymerize heparin by chemical (alkaline) elimination and (2) would adjust process conditions such that they result in manufacturing the same active
ingredient as Lovenox's enoxaparin. To manufacture enoxaparin, the process conditions could (but do not
necessarily need to) be the same as those used for Lovenox's enoxaparin. Insofar as an equivalent manufacturing
process for enoxaparin could be interpreted only to be one in which every step ofthe manufactuing process,
including process conditions, are identical to those of Aventis's manufacturing process for Lovenox, your request is
denied.
5 FDA classifies as therapeutically equivalent, and thus substitutable, those products that are (1) approved as safe
and effective, (2) pharmaceutically equivalent (which means, in part, drug products in identical dosage forms that
the identical active ingredient; and meet the identical compendial or other applicable
contain identical amounts of
1 (c)), (3) bioequivalent, (4)
identity, strength, quality, and purity, including potency (21 CFR 320.
standard of
adequately labeled, and (5) manufactured in compliance with Current Good Manufacturing Practice regulations.
See Orange Book, 30th Ed., at iv. The terms "therapeutically equivalent" and "substitutable" are used interchangeably in this response.
and the active ingredient has not been fully characterized. Based on our evaluation of all the
relevant data and other current relevant scientific information, our experience and expertise,
Agency precedent, and applicable law, we find that enoxaparinhas been adequately characterized for the purposes of approving naturally sourced generic enoxaparin; and we conclude that an
ANDA applicant for enoxaparin can demonstrate active ingredient sameness by meeting five
the active ingredient's "sameness."
which captues different aspects of
criteria,6 each of
In very general terms, the five criteria involve (1) the physical and chemical characteristics of
the source material and the method used to break up the polysacenoxaparn,(2) the nature of
charide chains into smaller fragments, (3) the nature and arrangement of components that
constitute enoxaparin, (4) certain laboratory measurements of anticoagulant activity, and (5)
the drug's effect in humans. The equivalence evaluation for these criteria
certain aspects of
the generic drug product's
generally is based upon qualitative and/or quantitative comparsons of
enoxaparn to multiple batches of Lovenox' s enoxaparin and takes into consideration the batchto-batch variability and sampling of Lovenox, and analytical test variability. The equivalence
the generic drug product's enoxaparin
evaluation demonstrates that the molecular diversity? of
and Lovenox's enoxaparn is equivalent, including with respect to the 1,6 anhydro ring structure
at the reducing ends of
between 15 percent and 25 percent of its poly( oligo )saccharides.
Equivalent molecular diversity demonstrates sameness for enoxaparin.8 These five criteria
together comprise a robust test that provides overlapping evidence by which an ANDA applicant
for enoxaparin can demonstrate active ingredient sameness for enoxaparin within the meaning of
the Act and FDA regulations.
This approach to determining active ingredient sameness for enoxaparin has been considered
extensively by various components ofthe Agency's Center for Drug Evaluation and Research
(eDER), including the Office of
Pharmaceutical Science (including the Office of
Generic Drugs,
Biotechnology Products), and the

Gastrointestinal and Coagulation Drug Products
New Drugs (including the Division of
Office of
Medical Imaging/Hematology Products).9 We made this decision after
and the Division of
carefully considering the petition (including supplements and comments), other correspondence,
relevant scientific publications regarding characterizations ofLMWHs, ANDAs for enoxaparin,
and other relevant information. As with all complex scientific issues, it is possible that with
the Office of
New Drug Quality Assessment, and the Office of
improvement in the understanding of the biological and clinical properties of enoxaparn and/or
advances in the analytical technologies that might be used to characterize enoxaparin, other
approaches might emerge to establish the sameness of enoxaparin.
6 The terms "criteria" and "criterion," as used in this response, refer to a particular demonstration or showing of a
specific aspect of active ingredient sameness.
7 See footnote 25.
8 As discussed in section I.B, it is well established that certin activities (anti-Xa activity and anti-lla activity)
explain, in significant part, the pharmacological activity for low molecular weight heparis (LMWHs) (including
enoxaparin). Other mechanisms of action also may be important for the pharmacological activity of LMWHs
(including enoxaparin). The five criteria ensure that generic enoxaparin wil have the same active ingredient
Loven
ox's enoxaparin (within the context 6fits variability) even though the contribution of
components as those of
each component has not been fully elucidated. Therefore, pharacological activity ofthe active ingredient of the
Lovenox can be expected to be the same.
generic enoxaparin and that of
9 As a result of CDER's reorganization, responsibility for reviewing the NDA for Lovenox was tranferred from
Medical ImagGastrointestinal and Coagulation Drug Products to the Division of
CDER's Division of
ing/Hematology Products.
In addition to the issues raised in your petition, we have also considered issues related to
the generic
immunogenicity. It is important that ANDA applicants assess the potential of
product to generate a greater immune response as compared to the RLD, Lovenox.

This response contains seven sections. Section I contains relevant background information.
Section II describes the statutory and regulatory framework for active ingredient sameness.
Section III discusses the five criteria that provide sufficient information to enable us to conclude
active ingredient sameness for enoxaparn. Section iv explains that our decision on enoxaparin
sameness is consistent with previous ANDA approval decisions. Section V summarzes the
relevant case law, which supports our decision. Section Vi addresses specific arguments and
information in your petition and related comments and explains why those arguments and
information do not preclude a finding of active ingredient sameness for enoxaparin. Finally,
section VII summarzes our conclusion.
DISCUSSION
I. BACKGROUND
A. Lovenox
FDA approved Aventis's new drug application (NDA) for Lovenox (enoxaparin sodium
injection, NDA 20-164) on March 29, 1993.10 Lovenox is indicated for:
. Prophylaxis of deep vein thrombosis (DVT) in abdominal surgery, hip replace-
ment surgery, knee replacement surgery, or medical patients with severely restricted mobilty during acute ilness
. Inpatient treatment of acute DVT with or without pulmonary embolism
. Outpatient treatment of acute DVT without pulmonary embolism

10 Upon approval of
Lovenox, Aventis received 5 years of marketing exclusivity and other patent listing and
marketing
the Act. Aventis subsequently received 3 years of
Lovenox labeling related to ST-segment elevation
necessar clinical studies in support of
certification benefits described in section 505 of

exclusivity for conducting
patent certifications
myocardial infarction (STEMI). This exclusivity expired on May 16, 2010. As a result of
the Act and notice of
submitted by ANA applicants under section 505(j)(2)(A)(vii) of
those certifications, Aventis
fied a lawsuit against those ANA applicants for infrgement of patents submitted by Aventis to FDA as required
under section 505(b) and (c). After a tral, the distrct court found that U.S. Patent No. 5,389,618 (the '618 patent)
and its replacement, U.S. Reissue Patent No. 38,743 (the '743 patent) were unenforceable on the grounds that
Aventis committed inequitable conduct before the United States Patent and Trademark Office (PTO) in failing to
disclose material information to the PTa. See Aventis Pharma SA. v. Amphastar Pharmaceuticals, Inc., 475 F.
Supp. 2nd 970 (C.D. CaL. Feb. 8, 2007). The United States Court of Appeals for the Federal Circuit affired that
determination. See Aventis Pharma SA. v. Amphastar Pharmaceuticals, Inc., 525 F.3d. 1334 (Fed. Cir. May 14,
2008). The Supreme Cour denied the petition for certiorari. See Aventis Pharma SA. v. AmphastarPharmaceuti27, 2009). ANAs referencing Lovenox were submitted to FDA before the
cals, Inc., et al. 129 S.Ct. 2053 (April
the Medicare Prescription Drug, Improvement, and Modernzation Act of2003 (Public Law 108-173)
passage of
(MMA). Therefore, l80-day exclusivity for drug products referencing Lovepox is governed by section
the MMA. In
of the MMA. See Section 1102(b)(l) of
the Act as in effect before passage
505(j)(5)(B)(iv) of
accordance with the applicable exclusivity provisions, any l80-day exclusivity for these products was triggered on
October 2, 2008, when the Federal Circuit issued the Mandate with respect to its May l4, 2008 decision affirng
the district court's finding that the '618 and '743 patents are unenforceable due to inequitable conduct. The 180-day
period expired on March 31, 2009.
. Prophylaxis of
unstable angina and non-Qwave myo-
ischemic complications of
cardial infarction (MI)

. Treatment of acute ST -segment elevation myocardial infarction (STEMI) man-
aged medically or with subsequent percutaneous coronary intervention (PCI).I i
Lovenox contains the active ingredient enoxaparin sodium and is par of a relatively new class of
anticoagulants called LMWHs. Enoxaparin is composed of a diverse mixture of many oligosach' 12
Chan'd
e cams.
B. Regulatory History of Heparin and Low Molecular Weight Heparins Such as
Lovenox
LMWHs (such as enoxaparin) are manufactured by depolymerizing heparin sodium (heparin)
polysaccharide chains into correspondingly shorter oligosaccharide chains.13 Accordingly, it is
helpful to briefly discuss the regulatory history of heparin before discussing that of LMWHs.
was the first widely used
anticoagulant and continues to be widely used in the United States. Heparin is prepared by
processes involving extraction from animal tissues (i.e., porcine intestinal mucosa or bovine
lungs ).14 Concerns over the potential risks posed by bovine spongiform encephalopathy (BSE)
Heparin, as an anticoagulant, has been on the market since 1939. It
have made porcine tissue the principal source of
heparin. Heparin products currently used in the
United States are of porcine origin. Heparin is biosynthesized in the pig gut, rather than through
the active ingredient, and its manufacturing process entails (1) purification
chemical synthesis of
of
the heparin to separate it from other tissue components and (2) clearance or inactivation of
potential pathogenic organisms.
Heparin is generally administered intravenously and remains one of the anticoagulants of choice
heparin can be
when a rapid anticoagulant effect is needed. The anticoagulant effects of
monitored by widely available laboratory tests, and an antidote is readily available to quickly
reverse the drug's anticoagulant effects. The approved heparin labeling includes general
indications and dosage and administration instructions that reflect the need for frequent monitorheparin, its risk-benefit
ing.I5 Because ofthe long-standing and widespread clinical use of
including heparin-
profie is well known. The drug is known to be associated with serious risks,
induced thrombocytopenia (HIT), which is a serious antibody-mediated reaction resulting from

irreversible aggregation of platelets. Because patients vary widely in their response to heparin,
heparin therapy is generally restricted to hospital and other settings where its dose can be
adjusted on a closely monitored patient. Currently, there are eleven approved NDAs and six
II Product Labeling for Lovenox (enoxaparin sodium injection), NDA 20-164; Revised December 23,2009.
12 See footnote 3.
polysaccharide chains into smaller oligosaccharide

fragments by chemical or enzymatic means. Because LMWH chains are shorter than the parent heparin chains, we
generally use in this response the term "oligosaccharides" in connection with LMWHs and the term "polysaccharides" in connection with heparin.
14 Mulloy, 8., Gray E., Barrowcliffe, T.W. (2000), "Characterization of
Un fractionated Heparin: Comparison of.
Materials From the Last 50 Years," Thromb Haemost 84:1052-1056.
15 See, e.g., PrOduct Labeling for Heparin Sodium Injection USP, NDA l7-037; Revised December 2007. (Carton
13 Depolymerization refersto the breaking up (or cleavage) of
and Immediate Container Labels for Heparin Sodium Injection USP, NDA 17-037; Revised October 2009).
approved ANDAs for heparin sodium injection, which are listed in FDA's Approved Drug
16
Products with Therapeutic Equivalence Evaluations (Orange Book).
LMWHs, as noted above, are manufactured through depolymerization ofheparn.17 Both

heparin and LMWHs act as anticoagulants by inactivating both factor Xa and factor IIa in the
18, 19 Other mechanisms may also account for the anticoagulant activity of
coagulation cascade.
tissue
factor pathway inhibitor (TFPI), which prevents the progression of the coagulation cascade.20 In
comparison to heparin, LMWHs possess a longer half-life, a more predictable anticoagulant
response, and reduced side effects of fatal adverse reactions related to thrombocytopenia.
Accordingly, LMWHs do not require frequent laboratory monitoring of coagulation parameters
and are used in a wider range of treatment settings, including outpatient settings.
these drugs. For example, heparin and LMWHs are known to promote the release of
To date we have approved four NDAs for LMWHs: Lovenox (enoxaparin sodium)(NDA 20164), Fragmin (dalteparin sodium) (NDA 20-287), Normiflo (ardeparin sodium) (NDA 20-227)
the manufacturer),21 and Innohep (tinzaparin sodium) (NDA
(withdrawn in 2001 at the request of
ingredients in these NDA-LMWH products are derived from different
20-484). The active
modes of depolymerization22 yielding drug products containing active ingredients with different
distributions of oligosaccharide sequences and different chemical modifications at the terminal
these oligosaccharide chains.23 These NDA-LMWH products are regarded as containing
ends of
are not substitutable. Today, more than 15 years after the
different active ingredients, and thus
16 The approved generic hepari products are not expected to pose any safety or effectiveness risks beyond those
already associated with their
respective RLDs. The recent severe anaphylactoid reactions reported following
intravenous heparin administration were not attbuted to the hepari active ingredient in approved products, but
rather to the presence of over-sulfated chondroitin sulfate '(OSCS), a synthetic contaminant introduced into the
hepari active ingredient. See Kishimoto, T.K. et aL. (2008), "Contaminated Heparn Associated with Adverse
Clinical Events and Activation of
the Contact System," New England Journal Medicine 358: 2457-2467; Guerrni,
M. et aL. (2008), "Oversulfated Chondroitin Sulfate is a Contaminant in Heparin Associated Clinical Events," Nature
this contamination, the u.s. Pharmacopeia (USP) has revised its

monograph for Heparin Sodium USP to incorporate, among othr things, a nuclear magnetic resonance (NMR) and
identity for heparin sodium
the standard of
a high performance liquid chromatography (HPLC) test as part of
Biotechnology 26: 669-675. As a result of
(October 1,2009). These NMR and HPLC chromatographic tests are also used to detect the presence of
the OSCS
containant. FDA expects all approved NDAs and ANAs for Heparin Sodium Injection USP to comply with the
updated USP standard to detect the presence ofOSCS.
17 Linhardt, R.J., Gunay, N.S. (1999), "Production and Chemical Processing of
Low Molecular Weight Heparin,"
Semin Thromb Hemost 25 S3:5-L6.
18 Factor Xa is a protein factor in the coagulation cascade that activates prothombin (factor II) into thrombin (factor
IIa). Thrombin is a factor in the coagulation cascade that converts fibrinogen to fibri monomers, which eventually
self-associate and are cross-lined durig clot formation (blood coagulation).
19 Lin, R., Hu, Z. (2000), "Hematologic Disorders," in Melmon and Merrelli's Clinical Pharmacology, Carrthers,
S.G. et ai., eds., 4th ed., New York: McGraw Hil, 737-797.
20 Hirsh, J., Warkentin, T.E., Shaughnessy, S.G., Anand, S.S., Halperin, J.L., Raschke, R., Granger, C., Ohman,
E.M., Dalen, J.E. (2001), "Hepari and Low-Molecular Weight Heparin: Mechanisms of Action, Pharmacokinetics,
Dosing, Monitoring, Effcacy, and Safety," Chest LI9:64S-94S.
21 67 FR 6264 (February ll, 2002).
22 We use in this response the term "depolymerization mode(s)" or "mode(s) of depolymerization" to refer to the
chemical ( or enzymatic) means by which heparin is cleaved into LMWH.
23 The termnal ends are the ends of a heparin or LMWH saccharide chain. Each chain contain a "reducing" and
"non-reducing" end.
first NDA approval of a LMWH product (i.e., Lovenox), FDA is approving the first ANDA for a
LMWH product (i.e., an ANDA for enoxaparin).14
c. Molecular Diversity25 of LMWH
heparin is important for understanding the molecular

linear polysaccharides that are variable in length,
consisting of disaccharde26 repeating units composed of glucosamine and uronic acid (either
iduronic or glucuronic acid) with the following linkage sequence: ((1-+4) a-D-glucosaminylKnowledge of
the molecular diversity of
diversity of LMWH. Heparin is a mixture of
(1-+4) -D-hexuronosYl)n (Figure 1). Thus, as explained more fully below, the molecular
diversity of heparin comes from (1) the polydispersit~7 of chain length and (2) the diversity of
disaccharide units and the corresponding distrbution of disaccharide unit sequences in the
polysaccharide chains.28
0' 0 0 0 0 0 0 0
5 01 oSOi OOSOi ~C00-2' OOSOi CO2 00SOi
~. :'. '.j" ." ".~ "0; .J
ac ac ac ac
I ~SOj __~::o I ~H ~c I ~ '-AC I ~H ~o I

Figure 1. Representative polysaccharide in heparin showing four disaccharide units. These repeating units are
composed of glucosamine and a uronic acid (either iduronic or glucuronic acid) with the linkage sequence: (( 1 ~4)
a-D-glucosaminyl- (l ~4) -D-hexuronosYl)n.
The
first dimension of
heparin is the polydisperse chain length of
the molecular diversity of
polysaccharides. These polysaccharides have molecular weights ranging from 5,000 to 40,000
daltons (Da) and an average
molecular weight of ~ 15,000 Da. The polydispersity of
heparin,
characterized by a ratio of weight average to number average molecular weight (Mw/Mn), varies
between approximately 1.1 and 1.6.19
heparin's molecular diversity derives from the diversity of disaccharide

and the corresponding distrbution of disaccharide unit sequences in the polysaccharide
The second dimension of

units
chains. The diversity of disaccharide units (constituent building blocks of polysaccharide
two different uronic acid components (iduronic or

chains) arises not only from the possibility of
glucuronic acid), but also from differential modification at four possible positions of the
the uronic acid and C3 and C6 of
the
disaccharide unit. For example, the second carbon (C2) of
24 We approved Sandoz Inc.'s (Sandoz) ANA (77-857) for enoxaparin. The agency has determned that Sandoz's
ANA meets all the requirements for ANA approval, including those for active ingredient sameness.
25 The term "molecular diversity" refers to the heterogeneity of oligosaccharides and is used throughout this
response for brevity.
26 A disaccharide is a carbohydrate composed of two monosaccharides.

27 Polydispersity is a measure of
the distribution of
molecular weights in a given polymer, and is defined as weight
average molecular weight divided by the number average molecular weight. As polymer chains approach uniform
chain length, polydispersity approaches unty.
28 Capila, I., Linhardt, RJ. (2002), "Hepari-Protein Interactions," Angew Chern Int Ed 4l:391-412.
29 Linhardt, R.I. (2003), "Hepari: Strcture and Activity," J Med Chern 46:1-14.
glucosamine can be O-sulfated. In addition, e2 of the glucosamine can be N -acetylated or Nheparn originates from the biosynthetic p,athway of
sulfated.30 The molecular diversity of
heparn biosynthesis.3 i
Heparn can be depolymerized into LMWHs which have approximately one-third the molecular
weight of the parent heparin. The reduced average molecular weishts of LMWHs vary between
3000 and 7000 Da, and the polydispersity ranges from 1.1 to 1.5.3 Because these LMWHs are
obtained from the diverse mixture of polysaccharides that comprise heparin, the molecular
diversity associated with these LMWHs also derives, like heparin, from the polydispersity of
chain lengths, the diversity of disaccharide units, and the corresponding distribution of disaccharide unit sequences in the oligosaccharide chains.
Unlike heparin, LMWHs may have unique chemical modifications (or "fingerprints") at the nonthe oligosaccharide chains, which provide an
reducing and reducing ends (or terminal ends) of
additional, third dimension of molecular diversity not present in heparin. These unique chemical
de polymerization used
the mode of
fingerprints at the terminal ends ofthe chains are a result of
to manufacture the LMWH.33 For instance, enoxaparin sodium is derived by esterification34 of
heparn derived from porcine intestinal mucosa to the corresponding heparin benzyl ester. The
intermediate heparn benzyl ester undergoes alkaline -elimination cleavage.35 This results in
heparn
selective cleavage between the uronic and glucosamine residues and fragmentation of
polysaccharides into smaller oligosaccharide fragments, with the majority of the components
the chain.36 On the other
heparin polysaccharides through deaminative
having a 4,5 delta (4,5)-uronate structure at the non-reducing end of

hand, in the case of dalteparin, depolymerization of
cleavage using nitrous acid results in oligosaccharide fragments with a 2,5-anydro-D-mannitol

structure at the reducing end ofthe chain.37 Because of differing depolymerization modes, the
four approved LMWHs (enoxaparin, tinzaparn, dalteparin, and ardeparn) differ in their terminal
ends as well as their distribution of disaccharide unit sequences in the oligosaccharide chains.38
30 To ilustrate, in Figure 1 above, in the first disaccharide unit, C2 ofiduronic acid is O-sulfated and C2 and C6 of
glucosamine are N-sulfated and O-sulfated, respectively; in the second disaccharide unit, iduronic acid is not
modified, and C2 and C6 of glucosamine are N-acetylated and O-sulfated, respectively.
31 Capila, i. et aL. 2002.
32 Linhardt 2003.
33 Linhardt 1999; European Pharmacopeia (EP), 5th ed., 2006, Enoxapari Sodium, at 3493-3494; EP, Daltepar
Sodium, at 3925-3926; EP, Tinaparin Sodium, at 2586; Fragmin (daltepari sodium) injection package insert;
Lovenox (enoxaparin sodium) injection package insert; Innohep (tinzapari sodium) injection package insert;
Particular LMW Hepariic Polysacchardes for the ProphyDebrie, R. (February l4, 1995), "Mixtues of
laxis/Treatment of Acute Thrombotic Events," U.S. Patent No. 5,389,618 (February 14, 1995).
34 Esterification, in this instance, is a process that modifies the carboxylate of the uronic acid strctures into the
corresponding benzyl ester.
35 Specifically, the approved product labeling for Lovenox states that "enoxapari sodium is obtained by alkaline
heparin benzyl ester derived from porcine intestinal mucosa."
(i.e., chemical) depolymerization of
36 Linhardt 1999; EP at 3493-3494; Lovenox injection package insert; Debrie 1995.
37 Linhardt 1999.
38 As noted, we use in this response the term "depolymerization mode(s)" or "mode(s) of depolymerization" to refer
to the chemical (or enzymatic) means by which heparin is cleaved into LMWH; however, we do not use the term to
refer to the specific process conditions (operating parameters such as temperature, pH and depolymerization time)
used to depolymerize heparin into LMWH. For example, the depolymerization mode used to manufactue
enoxaparin is "cleavage by alkaline -elimnation of
is "nitrous acid depolymerization of
the benzyl ester derivative of
hepari."
8
heparin," whereas for daltepari it
In sum, the molecular diversity of LMWHs comes from (1) the polydispersity of chain length,
(2) the diversity of the disaccharide units and corresponding distribution of disaccharide unit
modified terminal end disacchasequences in the oligosaccharide chains, and (3) the diversity of
ride units in the oligosaccharide chains.
II. THE AGENCY HAS BROAD DISCRETION UNDER THE ACT AND FDA
REGULATIONS TO DETERMINE SAMENESS OF ACTIVE INGREDIENTS
. A.. Summary of Legal Framework for ANDA Approval
The Drug Price Competition and Patent Term Restoration Act of 1984 (the Hatch-Waxman
the Act, which established the ANDA approval process.
Amendments) created section 5050) of
To obtain approval, an ANDA applicant is not required to submit clinical studies to establish the
safety and effectiveness of
the drug product. Instead, an ANDA applicant relies on the Agency's
safety
and effectiveness, an ANDA applicant must demonstrate, among other things, that the generic
the Act).39 In addition, an
drug product is bioequivalent to the RLD (section 5050)(2)(A)(iv) of
ANDA must contain suffcient information to show that the generic drug product has the same
use, route of administration, dosage form,
active ingredient(s), previously approved conditions of
strength, and (with certain exceptions) labeling as the RLD (sections 5050)(2)(A) and 0)(4) of
the Act). The Agency must approve the ANDA unless, among other things, the ANDA applicant
has provided insufficient evidence of the foregoing, or if the methods used in, or the facilities
the drug are inadequate to
and controls used for, the manufacture, processing, and packing of
assure and preserve its identity, strength, quality, and purity (section 5050)(4) ofthe Act).
previous finding that the RLD is safe and effective. To rely on FDA's previous finding of
The premise underlying the Hatch-Waxman Amendments is that drug products that are (1)
approved as safe and effective, (2) pharmaceutically equivalent,40 (3) bioequivalent, (4) adequately labeled, and (5) manufactured in compliance with Current Good Manufacturing Practice
regulations are therapeutically equivalent and can be substituted for each other with the "full
expectation that the substituted tiroduct will produce the same clinical effect and safety profile as
the prescribed (RLD) product." i
B. Summary of Statutory and Regulatory Provisions on Active Ingredient Sameness

the Act states that, for a single active ingredient drug product, an
the generic drug product
ANDA must contain information to show that the active ingredient42 of
Section 505(j)(2)(A)(ii)(I) of
39 Under the Act, "(a) drug shall be considered to be bioequivalent to a listed drug if. . . the rate and extent of
absorption of
the listed drug

the therapeutic ingredient under similar experimental conditions in
the drug do not show a significant difference from the rate and extent of
when administered at the same molar dose of
absorption of
either a single dose or multiple dses." See section 5050)(8)(B)(i); see also implementing regulations at 21 CFR
par 320.
40 See 21 CFR 320. i (c) (pharmaceutical equivalents means, in part, drug products in identical dosage forms that
the identical active ingredient and meet the identical compendial or other applicable
identity, strength, quality, and purity, including potency).
contain identical amounts of

standard of
41 See footnote 5.
42 FDA regulations (at 21 CFR 210.3(b)(7)) provide that "(aJctive ingredient means any component that is intended
to furnish pharmacological activity or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention
the Act, we must

approve an ANDA referencing a listed drug that has only one active ingredient unless the ANDA
contains insuffcient information to show that the active ingredient is the same as that of the
listed drug.
is the "same" as that of
the listed drug. Under section 505(j)(4)(C)(i) of
information that an ANDA

applicant must submit to demonstrate that the active ingredient in the generic drug product is the
same as the active ingredient in the RLD, nor do these provisions describe the type or amount of
information on which we may rely in determining whether the ANDA applicant has provided
sufficient information to show that the active ingredient is the same. Accordingly, Congress
of
These statutory provisions do not describe the type or amount
recognzed that we must have broad discretion with respect to the information we may consider
in making a finding on the "sameness" of an active ingredient. 43
Parallel FDA regulations implementing these statutory provisions (i.e., sections 505(j)(2)(A)(ii)
and (j)(4)(C)) can be found at 21 CFR 314.94(a)(5)(i) and 314.127(a)(3). FDA regulations also
provide that an ANDA is suitable for consideration and approval if the generic drug product is
the same as the RLD (21 CFR 314.92(a)(I)). Specifically, 314.92(a)(I) states that the term
"same as" means, among other things, "identical in active ingredient(s)." In the preamble to the
the Hatch-Waxman Amendments, we specifically rejected the
final rule implementing title I of
suggestion that we adopt a requirement that active ingredients "exhibit the same physical and
chemical characteristics, that no additional residues or impurities can result from the different
manufacture or synthesis process; and that the stereochemistry characteristics and solid state
forms ofthe drug have not been altered.,,44 Instead, we adopted a more flexible approach, stating
that we would "consider an active ingredient (in a generic drug product) to be the same as that of
the reference listed drug ifit meets.the same standards for identity.,,45 We further stated that, in
most cases, the standards for identity are described in the USP, although we might prescribe
"additional standards that are material to the ingredient's sameness.,,46 In the case of enoxaparin,
there is a USP monograph/7 and there are additional standards that are material to enoxaparn's
sameness.
As FDA's regulations and preamble reflect, and as you acknowledge (Petition at 21), we have
broad discretion in determining whether an ANDA applicant has submitted suffcient information upon which we can reasonably conclude that the generic drug product's active ingredient is,
the RLD.
as a matter oflaw, the "same" as that of
of disease, or to affect the strcture or any function of the body of man or other animals. The term includes those
components that may undergo chemical change in the manufacture of the drug product and be present in the drg
product in a modified form intended to furnish the specified activity or effect." FDA regulations (at 21 CFR
3l4.3(b)) also provide that "drug substance means an active ingredient that is intended to fuish pharmacological
activity or other direct effect in the diagnosis, cure, mitigation, treatment, or prevention of disease or to affectthe
strcture or any function of the human body, but does not include intermediates user dJ in the synthesis of such
ingredient. "
43 See generally Serono Laboratories, Inc. v. Shalala, 158 F.3d 1313 (D.C. Cir. 1998); see also discussion in section
V of this response.
44 See 57 FR 17950 at 17958-59 (April 28, 1992).
45 Id. at 17959.
46 rd.
47 See USP 32-NF 27 monograph for enoxaparin sodium (offcial on December 1,2009).
10
III. GENERIC ENOXAPARIN CAN CONTAIN THE SAME ACTIVE INGREDIENT

(ENOXAPARIN) AS LOVENOX WITHIN THE MEANING OF THE ACT AND
FDA REGULATIONS
As discussed above, an ANDA applicant for generic enoxaparin must provide sufficient
information to show that the generic drug product contains the "same" active ingredient (enoxaparin) as Lovenox. We have concluded, based on our evaluation of current data and other
current relevant scientific information and our scientific experience and expertise, that the
following five criteria (or standards for identity) together provide suffcient information to
conclude that generic enoxaparin has the "same" active ingredient as Lovenox:48
1. Equivalence of physicochemical properties

2. Equivalence of
heparin source material and mode of depolyierization
3. Equivalence in disaccharide building blocks, fragment mapping, and sequence of oli-
gosaccharide species
4. Equivalence in biological and biochemical assays
5. Equivalence of
in vivo pharacodynamic profile
These five criteria take into account the inherent molecular diversity associated with Lovenox's
the disacchaenoxaparin and address (1) the polydispersity of chain length, .(2) the diversity of
ride units and corresponding distribution of disaccharide unit sequences in the oligosaccharide
chains, and (3) the diversity of modified terminal end disaccharide units in the oligosaccharide
to-batch varability, which you acknowledge is
batchchains. Lovenox has some degree of
expected in any product derived from living organisms.49 The equivalence evaluation for these
criteria generally is based upon qualitative and/or quantitative comparisons ofthe generic drug
Lovenox's enoxaparin and takes into consideration
product's enoxaparn to multiple batches of
the batch-to-batch varability and sampling of Lovenox, and analytical test varability. The
equivalence evaluation for the above five criteria demonstrates that the molecular diversity ofthe
generic drug product's enoxaparn and Lovenox's enoxaparin wil be equivalent. Equivalent
molecular diversity demonstrates sameness for enoxaparin.50 Collectively, the five criteria are
designed to provide overlapping evidence upon which we can conclude that the generic drug
product's enoxaparn is the same as Lovenox' s enoxaparin.
We provide below an explanation of
these five criteria is important to establish
why each of
sameness of enoxaparin.
A. Criterion 1: Equivalence of Physicochemical Properties
The first criterion for demonstrating sameness ofenoxaparin is equivalence of physicochemical

properties, such as molecular weight distribution and overall chemical composition. Equivalence
48 By vire of satisfying the five criteria described in this section, the standards for identity described in the USP for
enoxaparin and additional standards material to the ingredient's sameness are met.
49 Aventis October 13,2004, Comments at 20 (acknowledging that, "(oJf course, it should be noted that some degree
of
variation is expected in any product derived from living organisms").
50 See footnote 8.
11
in these properties provides, for the most part, information on the broad characteristics of
enoxaparin and is thlls an important element in establishing active ingredient sameness.
this response, one element of enoxaparin's molecular diversity is the
As stated in section i.e of
polydispersity of the oligosaccharide chain lengths. Therefore, it is important that the generic
drug product's enoxaparin has a distribution of oligosaccharide chain lengths equivalent to that
of
Lovenox's enoxaparn.
The molecular weight distribution determination, which is generally performed using size
exclusion chromatography, provides information on the relative abundance of oligosaccharides
of different molecular weights that comprise enoxaparin. Testing using size exclusion chromatography constitutes an important element for demonstrating equivalency of the oligosaccharide
chain lengths, including their distribution and proportion. Due to the inherent low resolution of
conventional size exclusion chromatography, it is also important to conduct a complementary

analysis termed "chain mapping." Chain mapping involves using methodologies, such as CTASAX (cetyltrimethylammonium coated strong anion exchange) chromatography,5i MALDI-MS
(matrix-assisted laser desorption ionization mass spectrometry), GPC-ESI-MS (gel permeation
spray ionization mass spectroscopy), or RPIP-ESI-MS (reverse phase ion
chromatograph - electro
pair - electro
spray ionization mass spectroscopy)52 to provide a high resolution fingerprint profie
the molecular weight distribution using both conventional size exclusion chromatography and complementary high resolution
of oligosaccharide chain lengths. Demonstrating equivalence of
heparin in
chain mapping helps to ensure that the extent and the pattern of depolymerization of
Lovenox's enoxaparin.
the generic drug product's enoxaparn is equivalent to that of
In addition to these analyses, it is also important to demonstrate equivalence of

chemical composition of
the overall
the generic drug product's enoxaparin and Lovenox's enoxaparn. This
type of analysis captures broad aspects of
the aggregate mixture of oligo
saccharides that
comprise enoxaparn. For example, nuclear magnetic resonance (NMR) spectroscopy can be
used to provide "spectroscopic fingerprints" of several of enoxaparin' s characteristic strctures,
such as the epimerization state of the uronic acid structure (i.e., iduronic versus glucuronic acid),
and the ratio of sulfated and non-sulfated .64,5 -uronate structures at the non-reducing end of the
ultraviolet (UV) specific absorbance can also be used
oligosaccharide chains.53 Equivalency of
to demonstrate the presence of unique functional groups such as the .64,5 -uronate structure known
to be present in enoxaparin.54 Other criteria for showing physicochemical equivalence can also
be used that capture broad characteristics of enoxaparin, including, among other things, certain
tests and acceptance criteria described in the USP monograph for enoxaparin (e.g., equivalent
51 Mourier, P.A.J., Viskov, C. (2004), "Chromatographic Analysis and Sequencing Approach of
Heparin Oligosac-
charides Using Cetyltrmethylammonium Dynamically Coated Stationary Phases," Analytical Biochemistry

332:299-313.
52 Thanawiroon, C., Rice, K.G., Toida, T., Linardt, RJ. (2004), "Liquid Chromatography/Mass Spectrometr
Sequencing Approach for HigWy Sulfated Heparin Derived Oligosaccharides," The Journal of Biological Chemistry, 279: 2608-2615..
53 Chuang, W., Chrst, M.D., Rabenstein, D.L. (2001), "Determination of
Heparin- and
the Primary Strctures of
saccharides Using Band Selective Homonuclear-Decoupled Two Dimensional IH
Heparin Sulfate-Derived Oligo
NMR Experiments," Anal. Chern. 73:2310-2316.
54 See USP 32-NF 27 (offcial on December 1,2009).
12

l3C NMR spectra, quantitative determinations for sodium content, and ratio of sul-
fate/carboxylate).
physicochemical properties provides, for the most par, information on

equivalence of
In sum,
broad characteristics of enoxaparin, such as evidence of distribution of oligosaccharide chain
of
Lovenox's enoxaparin and is an important element for demonstratlengths equivalent to that
ing enoxaparin sameness. This criterion alone does not provide sufficient information (on the
composition and sequence of enoxaparin oligosaccharde chains) upon which we can conclude
enoxaparn sameness. Satisfaction of this criterion together with the remaining four criteria,
the generic drug product's enoxaparin and Lovenox's enoxaparn wil be equivalent and, therefore, provides sufficient information to conclude active ingredient (enoxaparin) sameness.
however, would demonstrate that the molecular diversity of
B. Criterion 2: Equivalence of Heparin Source Material and Mode of Depolymeriza-
tion
The second criterion for demonstrating the sameness of enoxaparin is equivalence 6fheparin
source material (i.e., heparin that is derived from porcine intestinal mucosa and that meets USP
monograph standards for Heparin Sodium USP) and mode of depolymerization (i.e., cleavage by
heparin). The importance ofthis criterion
alkaline -e1imination of
is best ilustrated by considering how heparin source material and mode of depolymerization
(1) the polydispersity of chain lengths, (2) the diversity of disaccharide units and corresponding distribution of
sequences of disaccharde units in the oligosaccharide chains, and (3) the diversity of modified
terminal end disaccharide units in the oligosaccharide chains (see section LC). Physicochemical
properties provide information on the polydispersity of chain lengths as previously described in
Criterion 1 (see section lILA), and once Criterion 1 has been met for a generic drug product's
enoxaparin, the other two dimensions of molecular diversity and consequently equivalence can
be addressed by the following two-part framework.
relate to the three important dimensions
of enoxaparin's molecular diversity:
First. based on a fundamental understanding of cleavage chemistry, there is essentially no

rearangement of the sequences of "natural" disaccharide building blocks in heparin during the
cleavage reaction of the parent heparin polysaccharide chains into enoxaparn oligosaccharide
chains. The resultant distribution of sequences of disaccharide units in enoxaparin is essentially
found "naturally" in heparin and (2) site(s) where the
both a function ofthe (1) sequences
cleavage reaction occurs in the parent heparin chains. Second, the new chemical structures
introduced at the terminal ends of the cleaved oligosaccharide chains of enoxaparn are a result
of the cleavage reaction by which the heparin polysaccharde chains are depolymerized to the
enoxaparn oligosaccharide chains (Figure 2). This two-part framework is described in more
detail below.
Part J of framework
If an ANDA applicant for enoxaparin shows that it uses the equivalent heparin source material
and mode of depolymerization as that used for Lovenox's enoxaparin, then we can conclude that
13
the resultant distribution of sequences between the two enoxaparin active ingredients wil be at
least similar.
With respect to the heparn source material, the molecular diversity of natural disaccharide
building block sequences within heparn results from its biosynthetic pathway. Thus, equivalent
heparin source material, for the purposes of manufacturing enoxaparin, wil be expected to have
natural disaccharide building block sequences (within the context
at least a similar distribution of
of its variability). Furthermore, if the equivalent mode of depolymerization is used, we can
expect the chemical selectivity of cleavage ofthe parent heparin polysaccharide chains for the
generic drug product's enoxaparin and Lovenox's enoxaparin to be at leastsimilar.55 However,
the chemical selectivity depends on both the cleavage chemistry (mode of depolymerization) and
the process conditions (operating parameters such as temperature, pH, depolymerization time)
56 For the chemical selectivity to be equivalent, such
used in the depolymerization process.
process conditions would need to be appropriately adjusted.
Part 2 of framework
the modified
disaccharide units at the terminal ends of the oligosaccharide chains that arise from the cleavage
57 Therefore, if
the mode of depolymerization used to cleave heparin chains is equivareaction.
The depolymerization process essentially determines the structural identities of
the cleavage patterns or site(s) of cleavage of
55 Chemical selectivity refers to, among other things, the biases of
heparin polysaccharide chains. The chemical selectivity of the mode of depolymerization determines the resultant
sequences of cleaved oligosaccharides. To ilustrate, a single hepari polysaccharide chain with a sequence of
ABCDEDA may be depolymerized through a process having a chemical selectivity with a bias to cleave adjacent to
two oligosaccharide fragments, AB and CDEDA. However, the single
the C disaccharide unit, and this results in
heparin polysaccharide chain with the same sequence ABCDEDA may be depolymerized through a process having a
different chemical selectivity with a bias to cleave adjacent to the D disaccharide unit, and ths results in three
oligosaccharide fragments, ABC, DE, and DA. Although there is no rearangement of
cleavage of
the sequences durg the
the hepari polysaccharide chain, the chemical selectivity of depolymerization affects the final
distribution of resuitant oligosaccharide sequences. As a result, if a given heparin material is depolymerized by two
different manufacturers with processes having the equivalent chemical selectivity, we can reasonably expect that the
resultant oligosaccharides wil have the equivalent distrbution of sequences. Conversely, if a given heparin material
is depolymerized by two different manufacturers with processes having different chemical selectivity, we can
reasonably expect that the resultant oligosaccharides wil have different distrbutions of sequences.
56 For example, enoxaparin and tinaparin are derived from heparin material isolated from porcine intestinal mucosa,
but are manufactued by different modes of depolymerization (i.e., chemical (alkaline) versus enzymatic -
elimination, respectively). These differig modes of depolymerization have vastly different chemical selectivities of
heparin polysaccharide chain cleavage. In the mode of depolymerization by enzymatic -elimination, cleavage
takes place exclusively in hepari polysaccharide chains at sites where the disaccharide unit has the 2-0sulfoiduronic acid strcture, whereas in the mode of depolymerization by chemical ( alkaline) -elimination,
cleavage occurs without preference for the presence or absence of a 2-0-sulfo group in the iduronic acid structure.
See Linhardt, R.I., Gunay, N.S. (1999), "Production and Chemical Processing of
Low Molecular Weight Hepari,"
Semin Thromb Hemost 25 S3: 10. If an ANA applicant chooses a depolymerization mode of alkaline -elimination
of the hepari benzyl ester that is equivalent to that used for Lovenox's enoxapari, then we can reasonably expect
the depolymerization process to be at least similar to that used for Lovenox's enoxapari
the chemical selectivity of
(i.e., relative cleavage at the 2-0-sulfoiduronic acid strctures versus unsulfated iduronic acid strctures would be at
least similar).
57 For example, daltepari is depolymerized by deaminative cleavage using nitrous acid. This mode of depolymerization has the effect of
introducing a modified disaccharide unit having a 2,5-anhydro-D-mannitol strcture at the
the oligosaccharide chains where cleavage occurs. On the other hand, enoxaparin is depolymerized
by alkaline (chemical) -elimination of the benzyl ester of heparin. This mode of depolymerization has the effect of
reducing end of
14
lent for the generic drug product's enoxaparin and Lovenox's enoxaparn, then we can expect the
saccharides to be at least similar.
modified disaccharide units at the terminal end of the oligo
Hlpari n
. Sequences of "natural" disaccharide units
/" ~
~ .~
~
Enapai n
Sequences of "natural" disaccharide units
Terminal "modified" disaccharide units
~ "Natural" disaccharide unit
o "Modified" disaccharide unit (non-reducing terminal end)
o "Modified" disaccharide unit (reducing terminal end)
"natural" disaccharide building blocks that derive

from heparin "natual" disaccharide building block sequences. Enoxaparin also contains strcturally modified
Figure 2. Enoxaparn consists of a distrbution of sequences of
termnal end disaccharide building blocks that arise from cleavage (depolymerization) of
the heparin oligosaccha-
rides.
In sum, we can reasonably conclude (provided there is equivalence of physicochemical properuses the equivalent heparin source
the ANDA applicant for enoxaparin shows that it
de polymerization as that used for Lovenox's enoxaparin, then
material and equivalent mode of
saccharides wil be at least similar with respect to both (1) the
the resultant mixture of oligo
"natural" sequences of disaccharide units in the oligosaccharde chains and (2)
distribution of
diversity of the modified disaccharide building blocks at the terminal ends of the oligosaccharide
chains. This provides important information for demonstration of enoxaparin sameness, but it is
not sufficient by itself to conclude enoxaparin sameness. Satisfaction of the remaining three
criteria in addition to the previous two criteria, however, would be suffcient to demonstrate that
ties) that if
introducing a modified disaccharide unit having a A 4,5 -uronate structure at the nonreducing end of the chain where
cleavage occurs. Therefore, the mode of depolymerization used to cleave heparin polysaccharide chains essentially
determines the tyes of structures (or chemical functionality) of the modified disaccharide unts present at the
the oligosaccharide chains in LMWHs.
terminal ends of
15
the molecular diversity ofthe generic drug product's enoxaparin and Lovenox's enoxaparin wil
be equivalent and, therefore, provides suffcient information to conclude that the generic drug
product's enoxaparin and Lovenox's enoxaparin are the same.
C. Criterion 3: Equivalence in Disaccharide Building Blocks, Fragment Mapping, and
Sequence of Oligosaccharide Species

The third criterion for demonstrating the sameness of enoxaparin is equivalence in disaccharide
building blocks, fragment mapping, and sequence of oligosaccharde species. If this third
criterion is met in addition to Criteria 1 and 2, then collectively the information provides
substantial supporting evidence that the distribution of sequences of disaccharide units in the
oligosaccharde chains and the structures of the modified disaccharide units at the terminal ends
of
the oligosaccharide chains of
the generic drug product's enoxaparin are equivalent to those in
1. Equivalence in disaccharide building blocks
Equivalence in disaccharide building blocks is established by demonstrating equivalence in the

composition (identity and quantitative levels) of disaccharide units (and other small oligosacchathe oligosaccharide chains in enoxaride units)58 which are the constituent building blocks of
parin.
Compositional analysis of these disaccharide building blocks can be achieved by exhaustive

digestion of enoxaparin with purified heparn digesting enzymes (heparinases I, II, III) and
nitrous acid, among other means, to yield the constituent disaccharide building blocks comprising enoxaparin. These individual disaccharide building blocks can then be separated and
quantified by a variety of approaches, including capilary electrophoresis (CE),59 reverse phase
high performance liquid chromatography (RP_HPLC),6o and strong anion exchange high
these disaccharde
building block units can be achieved using several techniques including comparison to structurally assigned disaccharide units in the literature,62 spectroscopic approaches such as mass
performance liquid chromatography (SAX_HPLC).61 Identification of
58 When we refer to the term disaccharide units in this section, the term also encompasses other small oligosaccha-
ride unts. Other small oligosaccharide units that are generated from this type of compositional analysis arise due to
the fact that while compositional analysis yields primarily disaccharide building block units, it can also yield (in
some instances) other small oligosaccharide units, such as trisaccharide units (which derive from enoxapari
oligosacchardes having an odd number of saccharide units) and tetrasaccharide units (which derive from the
inherent resistance of some tetrasaccharide units to fuher cleave into disaccharide units).
59 Sundarem, M., Qi, Y., Shriver, Z., Liu, D., Zhao, G., Venktaraman, G., Langer, R., Sasisekharan, R. (2003),
Low-Molecular Weight Heparis with Improved In Vivo Activity," Proc Nat! Acad Sci USA,
"Rational Design of
100 651-656.
60 Toyoda, H., Yamamoto, H., Ogino, N., Toida, T., Imanari, T. (1999), "Rapid and Sensitive Analysis in Heparin
Reversed-Phase Ion-Pair Chromatography on a 2 mm Porous Silica Gel Column," J.
and Heparin Sulfate of
Chromatography, A 830 197-20L
61 Muurier P., Viskov C., (June 2, 2005), "Method for Determining Specific Groups Constituting Heparins or Low
Molecular Weight Heparins," US Patent Application Publication US2005/01 19477 Al.
62 Mourier 2005. r
16

spectroscopy63 and NMR spectroscopy, 64 and chemical approaches such as analysis with
modifying reagents65 (e.g., sodium borohydride, nitrous acid) or modifying enzymes (e.g., 2-0sulphatase, 6-0-sulphatase, /l4,5 -glycuronidase). 66,67 These analyses can be used to identify the
structures of the disaccharide building block units including the sulfation and acetylation
substitution patterns at specific disaccharide hydroxyl or amino groups. They can also be used to
identify whether the disaccharide possesses, among other structures, a glucosamine, mannosamine, l,6 anydro ring,68 or galacturonic acid structure.
Such analysis can be used to determine the composition of "natural" disaccharide units69 that
comprise the oligosaccharide chains. It can also be used to determine the composition of
modified disaccharide units at the terminal ends of the oligosaccharide chains.7o, 71 When a
generic drug product's enoxaparin has been shown to have the same composition of disaccharide
units found in Lovenox's enoxaparin, we can conclude that the generic drug product's enoxaparin uses the same "natural" disaccharide units to assemble the distribution of sequences (of
disaccharide units) of oligosaccharide chains as Lovenox's enoxaparin. Moreover, from such
that the generic drug product's enoxaparin has the same
information we can also conclude
composition of modified disaccharide building blocks at the terminal ends of the oligosaccharide
chains as Lovenox's enoxaparin. This latter point is significant, as explained below.
Information derived from the analysis of the modified disaccharide building block units at the
paricular signifioligosaccharide chains in enoxaparin is of
non-reducing and reducing ends of
cance because, as stated in section III.B, the depolymerization process essentially determines the
structural identity of the modified disaccharide units at the terminal ends of the oligosaccharide
these modified disaccharide units at the terminal
chains. Moreover, the quantitative levels of
. ends of the oligosaccharide chains are quite sensitive to variations in the process conditions (or
operating parameters) - such as pH, temperature, and depolymerization time - that are used in
the benzyl ester
the given mode of depolymerization (i.e., cleavage by alkaline -elimination of
63 Saad, a.M., Leary, J.A. (2003), "Compositional Analysis and Quantification of
Heparin and Heparin Sulfate by
Electrospray Ionization Ion Trap Mass Spectrometry," Anal. Chern 75, 2985-2995.
64 Mourier 2005.
65 Mourier 2005.
66 Stringer, S.E., Balbant, S.K., Pye, D.A., Gallagher, J.T. (2003), "Heparin Sequencing," Glycobiology 13(2) 97103.
67 Myette, J.R., Shriver, Z., Kisiltepe, T., McLean, M.W., Venkataraman, G., Sasisekharan, R. (2002), "Molecular
Cloning of
the Heparin/Heparin Sulfate /14,5 Unsaturated GlyculOnidase From Flavobacterium Heparinum, Its
Recombinant Expression in Escherichia Coli, and Biochemical Determination ofIts Unique Substrate Specificity,"
Biochemistry, 41.7424-7434.
68 Equivalence in disaccharide building blocks together with equivalence in molecular weight distribution shows that
between 15 percent and 25

its poly(oligo)saccharide chains.
69 These "natural" disaccharide units comprising enoxaparin essentially derive from the disaccharide units that
generic enoxaparin contains the 1,6 anhydro ring structure at the reducing ends of
percent of
comprise the heparin source material (from porcine intestinal mucosa) used in the manufacture of enoxaparin.
Please refer to the discussion in section II.B.
70 Compositional analysis can also be use to identify and quantify the modified disaccharide units that may be
present in the middle ofthe oligosaccharide chains (as opposed to the terininal ends), such as those disaccharides
containing (among others) the galacturonic acid structure.
71 This is performed in conjunction with the determination of the ratio of sulfated and nonsulfated /14,5 -uronate
the oligosaccharide chains by, for example, NMR spectroscopy, as discussed in
strctures in the nonreducing end of
section lILA.
17
derivative of
heparin). For example, as you acknowledge in your petition (at 13), the content of
the oligosaccharide chains in Lovenox's

enoxaparin is quite sensitive to the process conditions that are used to cleave the benzyl ester of
heparin by alkaline -elimination. Therefore, an ANDA applicant would generally be expected
not only to use a mode of depolymerization equivalent to that used in the manufacture of
Lovenox's enoxaparn, but would also be expected to make appropriate adjustments in process
conditions to manufacture enoxaparin that has the same composition of these modified disaccharide building block units as Lovenox's enoxaparin.
the 1,6 anhydro ring structure at the reducing ends of
When an ANDA applicant makes appropriate adjustments in the depolymerization process

conditions to manufacture a generic drug product containing enoxaparn with th same composimodified disaccharide building block units as Lovenox's enoxaparin, the fact that the
tion of
modified building blocks are the same provides evidence that the adjusted process conditions
the
have a chemical selectivity of cleavage that is equivalent to the chemical selectivity of
depolymerization process used to manufacture Lovenox's enoxaparin. Thus, the analysis of
these modified terminal disaccharide building block units serves as a sensitive surrogate marker
of
the underlying chemistry and, therefore, the chemical selectivity of depolymerization that is
used in producing enoxaparin.
provides confirmation of the discriminatory
power of disaccharide building block unit compositional analysis to detect subtle differences in
the molecular diversity ofthe oligosaccharides comprising enoxaparn.72 Therefore, such
equivalence of disaccharide building block compositional analysis provides important evidence
that the generic drug product's enoxaparin is the "same" as Lovenox's enoxaparin.
Further, our review of relevant scientific information
2. Equivalence in fragment mapping
As discussed above, equivalence in the composition of disaccharide building block units in

conjunction with eriteria 1 and 2 described in sections lILA and IILB provides important
evidence for determining whether the generic drug product's enoxaparn is the "same" as
Lovenox's enoxaparin. However, it is important to conduct additional analyses to confirm that

the distribution of sequences of disaccharide building block units in the oligosaccharide chains of
the generic drug product's enoxaparin is the same as that of Lovenox' s enoxaparin. This
additional evidence of "sameness" of the distribution of sequences of disaccharide building block
units in the oligosaccharide chains can be obtained by fragment mapping as explained below.
72 This conclusion regarding the discriminatory power of disaccharide building block unit compositional analysis is
based on, among other things, data submitted by an ANAapplicant on development batches of drug product
containing "enoxaparin-like materiaL." The development batches of enoxaparin-like material were manufactued
using the equivalent heparin source material and mode of depolymerization as were used for the ANA registration
batches, but they were manufactured under different process conditions. The enoxaparin-like material demonstrated
Loven
ox's enoxaparin based on compliance with then-proposed USP standards for
importt characteristics of
a activity, and anti-Xa/anti-Ila ratio.
enoxaparin, including molecular weight distrbution, anti-Xa activity, anti-II
(The USP 32 NF 27 monograph for enoxapari became offcial on 12/01/09.) Although the enoxaparin-like material
was similar to Lovenox's enoxaparin, differences in the molecular diversity of oligosaccharides were readily
identified based upon discemable differences in the quantitative levels of disaccharide building blocks units
(particularly in the modified disaccharide building block units) relative to those present in Lovenox's enoxaparin.
18
Fragment mapping involves only a partial digestion of enoxaparin with heparinase enzymes
saccharides (as opposed to a full digestion
(e.g., heparinase I, among other enzymes) into oligo
these oligosaccharide fragments using
into disaccharide building blocks), followed by analysis of
73, 74 Analogous to a trtic map
methods such as RPLC-HPLC or SAX-HPLC (among others).
saccharides represents a
for proteins, the fragment map of partially digested enoxaparin oligo
signature of recurrng oligosaccharide sequences unique to enoxaparn, and thus provides global
information on sequences of oligo
saccharides within the enoxaparin strcture. When a generic
drug product's enoxaparin has been shown to have the same oligosaccharide fragment mapping
profile as Lovenox's enoxaparin, this information provides significant evidence that the generic
drug product's enoxaparin possesses the same recurrng global oligosaccharide sequence
segments. as those in Lovenox' s enoxaparin.
In addition, fragment mapping provides important information regarding both the sequence
structure of oligosaccharides present in the heparn source material and information related to the
chemical selectivity of the depolymerization process used in producing enoxaparin from heparin.
This is because, as noted in section IILB, the distribution of sequences of disaccharide units in
enoxaparin is both a function of the sequences present in the heparin source material and the
site(s) where the cleavage occurs in the parent heparin chains. Development data from an
ANDA applicant for enoxaparin also show that the profies of recurrng global oligosaccharide
sequence segments, as assessed by their oligosaccharide fragment mapping profies, are quite
sensitive to variations in the process conditions, which, as discussed, can affect the chemical
selectivity of cleavage of the heparin source materiaL.
the discriminatory
to detect subtle differences in the molecular
power of oligosaccharide fragment mapping
diversity of oligosaccharides comprising enoxaparin.75 Therefore, equivalence of fragment
mapping provides further corroborative evidence that the generic drug product's enoxaparin is
relevant scientific information provides confirmation of
Further, our review of
the "same" as Lovenox' s enoxaparin.

3. Equivalence in sequence of oligosaccharide species
As discussed above, equivalence in disaccharide building block units and equivalence of

fragment mapping, in conjunction with eriteria 1 and 2 described in sections lILA and IILB,
provides important evidence for determining whether the generic drug product's enoxaparin is
the "same" as Lovenox's enoxaparin. Additional information confirms through direct sequencing of oligosaccharides that the distribution of sequences of disaccharde building block units in
the oligosaccharide chains of the generic drug product's enoxaparin is the same as that of
73 Linhardt, R.J., Rice, K.O., Kim, Y.S., Lohse, D.L., Wang, H.M., Loganathan, D. (1988), "Mapping and
Quantification of
Heparin," Biochem. J. 254781-787.
the Major Oligosaccharide Components of
74 Chuang, W.L., McAllister, H., Rabenstein, D.L. (2001) "Chromatographic Methods for Product-Profie Analysis
and Isolation of
Oligo
saccharides Produced by Heparinase-Catalyzed Depolymerization of
Heparin," Journal of
Chromatography, A 932 65-74.

75 Maddineni, J., Walenga, lM., Jeske, W.P., Hoppensteadt, D.A., Fareed, J., Wah, R., Bick, R.L. (2006), "Product
Commercially Available Low-Molecular-Weight Heparis and their Generic Versions: Therapeutic

Individuality of
Implications, Clinical and Applied," Thrombosis/Hemostasis 12:267-276.
19
Recent advances in structural analysis of carbohydrates have made possible the direct sequencing
of oligosaccharide chains from enoxaparin. For example, this can be done through propertyencoded nomenclature (PEN) in conjunction with MALDI-MS,16, 77 by iterative chemical and
saccharides in conjunction with analysis by

polyacrylamide gel electrophoresis,78 or by enzymatic digestion in conjunction with NMR
spectroscopy.79 When a comparable subset of oligosaccharides from both a generic drug
product's enoxaparin and Lovenox's enoxaparin is isolated and shown to possess the same
sequence, this information provides further corroborative evidence that the generic drug
product's enoxaparin possesses the same distribution of oligosaccharide sequences as Lovenox's
enoxapann.
enzymatic digestion of fluorescent tagged oligo
Data from an ANDA applicant also show that the resultant sequenced oligosaccharides in
enoxaparin are quite sensitive to variation in the process conditions used, which, as discussed,
the heparin source materiaL80 This is particucan affect the chemical selectivity of cleavage of
the result of
being
saccharides, which, by virte of
larly true for the subset of shorter chain oligo
the most cleavage reactions of the heparin oligosaccharde chains, are those oligosaccharides
whose sequence identities are most dependent on the chemical selectivity of depolymerization
that is used to produce enoxaparin from heparn.81
As noted in section III.B, the distribution of sequences of disaccharide units in enoxaparin is a

the (1) sequences found "naturally" in heparn and (2) site(s) where the cleavage
function of
reaction occurs in the parent heparn chains. If there is equivalence in physicochemical proper-
ties, heparin source material, and mode of depolymerization together with this sensitive marker
of equivalent chemical selectivity (i.e., based upon data showing equivalence of disaccharide
compositional analysis, fragment mapping, and sequences of short chain oligosaccharides), this
provides evidence that the manufactung process for generic enoxaparn wil cleave
information
the heparin polysaccharide chains at sites equivalent to those for Lovenox's enoxaparin.
76 Venktaraman, G., Shrver Z., Raman, R., Sasisekharan, R. (1999), "Sequencing Complex Polysaccharides,"
Science 286:537-542.
77 Shrver, Z., Raman, R., Venkataraman, G., Turnbull, KJ., Toida, T., Linardt, R., Bieman, K., Sasikharan, R.
(2000), "Sequencing of 3-0 Sulfate Containing Hepari Decasaccharides with Partial Antithrombin II Binding
Site," Proc. Nat!. Acad. Sci. USA 97:10359-10364.

78 Turnbull, lE., Hopwood, ll, Gallagher, J.T. (1999), "A Strategy for Rapid Sequencing of
Heparin Sulfate and
Hepari Sacchardes," Proc. Natl. Acad. Sci. USA 96:2698-2703.

79 Yamada, S., Sakamoto, K., Tsuda, H., Yoshida, K., Sugiura, M., Sugahara, K. (1999)," Strctual Studies of
Octasaccharides Derived from the Low-Sulfated Repeating Disaccharide Region and Octasaccliaride Serine,"
Biochemistry 38 838-847.
80This conclusion is based on, among other thigs, data submitted by an ANA applicant on development batches of
drug product containng "enoxaparin-like material" as described in footnote 72. Although the enoxaparin-like
material in development was similar to Lovenox's enoxapari based upon the then-proposed USP standards for
molecular weight distribution, anti-Xa activity, anti-lla activity, and anti-Xa/anti-lla ratio,
enoxapar, including
discernable differences in the sequences of short oligosaccharides were identified. This information demonstrates
that the sequences of short oligosaccharides are quite sensitive to variations in the process conditions (or operating
parameters) used during depolymerization.
81 Ifthis condition is met (in conjunction with the other criteria for sameness), we can reasonably conclude that the
higher order oligosaccharide chains in the generic drug product's enoxapari wil be the same as those
higher order oligosaccharide chains have sequence
the cleavage process because they result from fewer
identities that are less dependent on the chemical selectivity of
sequences of
in Lovenox's enoxaparin. This is because the sequences of
cleavage reactions.
20
Docket No. FDA-2003-P-02.73
Therefore, we would expect the active ingredient in the generic enoxaparin to be the same as
Lovenox's enoxaparin with respect to the distribution of sequences of disaccharide units in the
oligosaccharide chains.
relevant scientific information provides
confirmation of the discriminatory power of oligosaccharide sequencing to detect subtle
differences in the molecular diversity of enoxaparin. 82 Therefore, equivalence of oligosaccharde
sequences provides further corroborative evidence that the generic drug product's enoxaparin is
Further, consistent with these conclusions, our review of
the "same" as Lovenox' s enoxaparin.
In sum, when the first two criteria described in sections lILA and III.B are met, the third criterion
_ equivalence of disaccharde building blocks, fragment mapping, and sequencing of oligosaccharde species - provides crucial evidence towards demonstrating that the molecular diversity of
the generic drug product's enoxaparin and Lovenox's enoxaparin wil be equivalent and,
therefore, provides important information to help conclude that the generc drug product's
enoxaparin is the same as Lovenox' s enoxaparn.
D. Criterion 4: Equivalence in Biological and Biochemical Assays
The fourth criterion for establishing sameness of enoxaparin is equivalence of in vitro biological
and biochemical assay results. Although the first three criteria together provide crucial evidence
of equivalent molecular diversity, this fourth criterion provides additional important evidence of
active ingredient sameness based on the biological and biochemical properties of enoxaparin.
biological assays, it is important to demonstrate that the
generic drug product's enoxaparin is equivalent to Lovenox's enoxaparin with respect to in vitro
biological assays fr relevant markers of anticoagulant activity, such as measurements based on
(among other things) aPTT (activated parial thromboplastin time) and Heptest prolongation
time.
To meet the criterion of equivalence of
biochemical assays, it is important to demonstrate that

the generic drug product's enoxaparin is equivalent to Lovenox's enoxaparin with respect to
factor Xa inhibition (anti-Xa) and factor IIa inhibition (anti-IIa). This biomeasurements of
chemical inhibitory effect on factor IIa and factor Xa in the coagulation cascade accounts for the
To meet the criterion of equivalence of
most thoroughly understood pharacological basis by which heparins and LMWHs (including
enoxaparin) function as anticoagulants.

AT-III binding pentasaccharide sequence motif that is present in heparin and LMWHs. This particular protein
The molecular basis for factor Xa inhibition (anti-Xa) is mediated by an
heparin polysaccharide chains
binding motif, which is present in approximately 30 percent of
and 15 to 25 percent ofLMWH oligosaccharde chains, has a high affnity for AT-III, an
82 This conclusion regarding the discriminatory power of oligosaccharide sequencing is based on, among other
things, data submitted by an ANA applicant on so-called "generic enoxaparins" marketed in other foreign
countries. Although the enoxaparin-like material was simlar to Lovenox's enoxapari, including molecular weight
distribution, anti-Xa activity, anti-lla activity, and anti-Xa/anti-lla ratio, differences in the molecular diversity of
oligosaccharides were readly identified based upon discemable differences in the subset of shorter oligosaccharide
sequences relative to those present in Lovenox's enoxaparin.
21
endogenous serpin inhibitor. Binding of this pentasaccharide sequence motif to AT-III results in
factor Xa in
an activated AT-III-polysaccharde complex that inhibits the proteolytic activity of
the coagulation cascade. Similarly, the molecular basis for factor I1a inhibition (anti-IIa) is
mediated by this particular AT-III binding pentasaccharide sequence. However, in this instance,
the AT-III -pol ysaccharide complex wil inhibit factor IIa only if the AT-III binding pentasaccharide sequence
is present in a polysaccharide chain at least 18 saccharide units in length.83
heparin polysaccharide chains are longer than 18 saccharide units,

Because a large proportion of
this results in heparn having an anti-Xa/anti-lla ratio of approximately 1. By contrast, because
LMWHs (including enoxaparn) are derived through controlled cleavage of heparin polysaccharide chains, most LMWH oligosaccharde chains have less than 18 saccharide units, and this
results in LMWHs having anti-Xa/nti-lla ratios that are greater than 1. Differences in anti-Xa
and anti-lla activities among the various approved LMWHs (which are readily measured using
standard amidolytic chromogenic assays)84 can be used to differentiate enoxaparin from other
LMWH products marketed in the United States. Accordingly, a demonstration of equivalence in
anti-Xa activity, anti-lla activity, and anti-Xa/anti-lla ratio between the generic drug product's
enoxaparin and Lovenox's enoxaparin provides important evidence of equivalence ofbiochemical characteristics - which, at a minimum, is responsible for an important and well-established
mechanism of action that explains, in significant part, the pharmacological activity for LMWHs
(including enoxaparin).
In sum, equivalence in biological and biochemical assay results provides important evidence of
enoxaparin sameness. This criterion by itself is not sufficient to conclude enoxaparin sameness.
the first four criteria together with the remaining criterion described below,
Satisfaction of
however, would be sufficient to demonstrate sameness of enoxaparin.
E. Criterion 5: Equivalence of In Vivo Pharmacodynamic Profie
The fifth criterion for establishing sameness of enoxaparin is equivalence of in vivo pharmacodynamic profile. The comparison of in vivo pharacodynamic profies is based upon measin vivo anti-Xa and anti-lla profiles. It is well established that the different
urements of
LMWH products approved in the United States have different pharmacodynamic profiles based
on their in vivo anti-Xa and anti-lla profiles.85 These differing pharmacodynamic profiles
might be due in part to differences in anti-Xa/anti-lla rati086 or molecular weight distribution of
oligosaccharde chains,87 among other reasons.88 Therefore, it is important that an ANDA

83 Hirsh J., Warkentin T.E., Shaughnessy S.G., Anand S.S., Halperin J.L., Raschke R., Granger C., Ohman E.M.,
Dalen J.E. (2001), "Hepari and Low-Molecular Weight Heparin: Mechanisms of Action, Pharacokinetics,
Dosing, Monitoring, Effcacy, and Safety," Chest 119 64S"94S.
84 See USP 32-NF 27 (official on December 1,2009).

85 Erickson, B.I., Soderberg, K., Widlund, L., Wandeli, B., Tengbom, L., Risberg, B. (1995), "A Comparative Study
Three Low-Molecular Weight Heparins (LMWH) and Unfractionated Hepari (UH) in Healthy Volunteers,"
Thrombosis and Haemostasis 73, 398-401.
86 Administration of
products with differing anti-Xa/anti-lla ratios would result in differences in in vivo pharacoa and anti-lla activities.
anti-X
dynamic profies, as these are assessed based on the in vivo profies of
87 This phenomenon derives from the fact that higher molecular weight poly( oligo )saccharide species are cleared
of
from the circulation more rapidly than the lower molecular weight poly(oligo)saccharide species. This in vivo
accumulation oflower molecular weight poly(oligo)saccharide chains (which tend to have lower anti-lla activity)
may result in differences in circulating anti-Xa/anti-lla ratios. See Hirsh, 1., Warkentin, T.E., Shaughnessy, S.G.,
Anand, S.S., Halperin, J.L., Raschke, R., Granger, C., Ohman, E.M., Dalen, J.E. (200l), "Heparin and Low-
22
applicant meet this fifth criterion to provide important evidence of active ingredient sameness.
In sum, we conclude, based on our scientific experience and expertise and current relevant
scientific evidence, that if an ANDA applicant meets each of the five criteria described in section
III, this robust showing enables us to conclude that the molecular diversity of the generic drug
product's enoxaparin and Lovenox's enoxaparin wil be equivalent and, therefore, provides
sufficient information to conclude that the generic drug product's enoxaparin is the same as
As with all complex scientific issues, it is possible that, with improvement in the understanding
of
the biological and clinical properties of enoxaparin and/or advances in the analytical technologies that might be used to characterize enoxaparin, other approaches might emerge to
establish the sameness of enoxaparin. Curently, however, the five sameness criteria specified
above constitute adequate standards for identity for establishing enoxaparin sameness.
iv. OUR APPROACH TO DETERMINING SAMENESS OF ENOXAPARIN is

CONSISTENT WITH AGENCY PRECEDENT
Our approach to determining the sameness of enoxaparin is consistent with our previous ANDA
are
approval decisions for other generic drug products containing active ingredients that
heterogeneous polysaccharides. Specifically, our conclusions of active ingredient sameness for
generic heparin and hetastarch are based on the relevant scientific information and our knowlthose active ingredients. As discussed below, to demonstrate active ingredient sameness
edge of
the Act does not require ANDA applicants to (1) completely characterize all the different
polysaccharides of enoxaparin by isolating, purifying, and sequencing each of its unique
polysaccharide chains and determining their relative abundance, (2) use the same manufacturing
process as that used for the RLD, or (3) conduct clinical studies to demonstrate equivalent safety
and effectiveness.
A. Our Approach to Determining Active Ingredient Sameness for Enoxaparin is

Consistent with Our Determination of Active Ingredient Sameness for Heparin
As previously noted, there are currently six approved ANDAs for heparin sodium injection,
which are listed in the Orange Book. These approved generic heparin products are considered
them
therapeutically equivalent to (and substitutable with) their respective RLDs, and two of
have been marketed for decades. The criteria for establishing active ingredient sameness were
Molecular Weight Heparin: Mechanisms of Action, Pharmacokinetics, Dosing, Monitoring, Effcacy, and Safety,"
Chest 1 i 9 64S-94S.
88 For example, enoxaparin (derived by chemical -elimination cleavage) and nadroparin (derived from deaminative
cleavage) have similar molecular weight distributions and ratio ofanti-Xa/anti-lla activities, but have different
pharmacodynamic profies (see, e.g., EP 5th. Ed. (2007) (5.6) Nadroparin Calcium, at 2075-2077; Collgnon, F.,
the
Fryman, A., Caplain, H., Ozoux, M.L., Roux, Y.L., Bouthier, J., Thebault, U. (1995), "Comparison of
Pharmacokinetic Profies of
the Three Low Molecular Mass Heparins - Dalteparin, Enoxaparin, and NadroparinThromboembolism)," Thrombosis
AdministeredSubcutaneously in Healthy Volunteers (Doses for Prevention of
and Haemostasis, 73(4) 630-640; Stiekema, J.C,J., van Griensven, J.M.T., Dinther, T.G.V., Cohen, AF. (1993), "A
Three Low Molecular Weight Heparins and Glycosaminoglythe Anti-Clotting of
Cross-Over Comparison of
curonan," British Journal of
Clinical Pharmacology, 36 51-56). .

23
based on the USP monograph for heparin sodium in place at the time of ANDA approvaL. 89 As a
A applicants showed
general matter, to demonstrate active ingredient sameness for heparin, AND
that the generic drug product's active ingredient (heparin) (1) was isolated from the same animal
the RLD (e.g., porcine intestinal mucosa) and (2) had appropriate potency
tissue source as that of
based on in vitro coagulation and anti-factor Xa activities.
Despite the relative simplicity of these criteria for demonstrating sameness, there has been no
evidence of significant risks related to safety or effectiveness with the approved generic heparin
products beyond those already associated with their respective RLDs. Particularly with respect
to safety, the recent severe anaphylactoid reactions reported following intravenous heparn
administration were not attributed to the heparin active ingredient, but to the presence of oses,
a synthetic contaminant introduced into the heparin active ingredient.9o Thus, based on relevant
heparin at the time of ANDA approval, the criteria
scientific information and our knowledge of
provided suffcient information to conclude that the generic drug product's heparin was the same
as the RLD' s heparin.
Compared to heparin, enoxaparin has greater molecular diversity in its oligosaccharides because
the additional depolymerization step in the manufacturing process. We conclude, based on
of
relevant scientific data and information and our scientific experience and expertise, that ANDA
applicants for enoxaparin can demonstrate active ingredient sameness to Lovenox's enoxaparin
by meeting the five criteria described in section III, and these criteria take into account the
molecular diversity forenoxaparin. We conclude that such a showing
additional degree of
demonstrates that the molecular diversity of the generic drug product's enoxaparn and
Lovenox's enoxaparin wil be equivalent and, therefore, provides sufficient information to
conclude that the generic drug product's enoxaparin is the same as Lovenox's enoxaparin. As
with heparin, we conclude that it is not necessary to completely characterize all the different
polysaccharide sequences, nor is it necessary to use the same manufacturing process as that used
for the RLD or to conduct clinical trals to demonstrate equivalent safety or effectiveness.
B. Our Approach to Determining Active Ingredient Sameness for Enoxaparin is

Consistent with Our Determiation of Active Ingredient Sameness for Hetastarch
Hetastarch (hydroxyethyl starch) is widely used as a plasma expander and in many ways is
chemically analogous to enoxaparin. Hetastarch is a mixture of polysaccharides derived through
chemical modification (hydroxyethylation with ethylene oxide) and chemical cleavage (acidhighly heterogeneous amylopectin polysaccharides (a plantcatalyzed depolymerization) of
derived natural product).91 Like enoxaparin, hetastarch consists of a distribution ofpolysacchaof repeating units (chemically
rides that differ in length as well as in composition and sequence
the standard
for identification applicable tests and acceptance criteria based upon (1) NMR, (2) HPLC chromatography, (3) antifactor Xa to anti-factor IIa ratio, and (4) presence for sodium. The USP monograph has also included quality tests
and limits for assay, inorganic, organic impurities, residual solvents, pH, bacterial endotoxins, sterility and loss on
drying. FDA expects all approved NDAs andANAs for Heparin Sodium Injection USP to comply with the
updated USP standards.
89 USP has revised its monograph (October 1,2009) for Hepar Sodium USP to incorporate as part of
90 See footnote 16.
91 Ferber, H.P., Nitsch, E., Forster, H. (1985), "Studies on Hydroxylethyl Starch Part II," Arzneim- Forsch/Drug
Res. 35 (I) 3 615-622.
24
heterogeneous modified monosaccharide units). On the other hand, unlike enoxaparin, which is
composed oflinear chain oligosaccharides, hetastarch polysacchardes possess chain branching.
modified
Therefore, the polysaccharide chains in hetastarch differ not only in their sequence of
monosaccharide units but also in chain branching.
We have approved four ANDAs for hetastarch, which are listed in the Orange Book. To
demonstrate active ingredient sameness for hetastarch, ANDA applicants: (1) demonstrated
equivalence of physicochemical properties, including molecular weight distribution of polysaccharide chains and overall chemical composition (e.g., extent of hydroxyethylation) and (2) used
the equivalent source material as the RLD (i.e., amylopectin polysaccharides derived from
plants), as well as an equivalent mode of chemical modification (i.e., hydroxyethylation with
ethylene oxide) and depolymerization (acid-catalyzed depolymerization).92 When comparing the
distributions, we took into account the lot-to-lot variation ofthe RLD, noting
that hetastarch is "extremely polydisperse.,,93 Based on relevant scientific infornation and our
knowledge ofhetastarch, these criteria provided suffcient information to conclude that the
generic drug product's hetastarch was the same as the RLD's hetastarch.
molecular weight
To demoIlstrate enoxaparin sameness, ANDA applicants meet not only criteria analogous to
those for determining sameness for hetastarch (including equivalence of physicochemical
properties, equivalence of source material, and mode of depolymerization), but also additional
criteria (i.e., equivalence of disaccharide building block units, sequence of oligosaccharide
species, fragment mapping, in vitro biochemical and biological assays, and in vivo pharmacodynamic parameters). As with hetastarch, we conclude that it is not necessary to completely
the different polysaccharide sequences, nor is it necessary to use the same
characterize all of
manufacturing process as that used for the RLD or to conduct clinical trals to demonstrate
equivalent safety or effectiveness.
In sum, our active ingredient sameness conclusion for enoxaparin is consistent with those for
other generic drug products containing active ingredients that are heterogeneous polysaccharde
mixtures. Our conclusion for enoxaparin (like those for heparin and hetastarch) is based on
relevant scientific information and our knowledge of the active ingredient.
V. OUR APPROACH TO DETERMINING ACTIVE INGREDIENT SAMENESS

COMPORTS WITH CASE LAW
The U.S. Cour of Appeals for the District of
Columbia's decision in Serono Laboratories, Inc. v.
Shalala, 158 F.3d 1313 (D.C. Cir. 1998), supports our approach to determining sameness of
enoxaparn. In Serono, the eourt of Appeals squarely addressed the issue of active ingredient
of
the Act and FDA regulations.
sameness within the meaning
92 Letter dated July 25, 1996, to Richard 1. Meader, McGaw, Inc., from Kathr C. Zoon and Janet Woodcock,
FDA, in response to Citizen Petition No. 96P-0024/CPl, at 2-3. This citizen petition was originally assigned docket
FDA's transition to its new
number 96P-0024/CPL. The number was changed to FDA-1996-P-000l as a result of
docketing system (Regulations.gov) in January 2008.
93Id.
25
Serono involved a legal challenge to our approval of a generic version of Pergonal, a menotropins product used to treat infertility. This product contains two active ingredients: follclestimulating hormone (FSH) and luteinizing hormone (LH). As the cour noted, we concluded
that to be the same, the generic drug product's active ingredients and Pergonals active ingredito-batch
batchents were expected to have the same primary structure, potency, and degree of
the active ingredients included natural variations known as microheterogeneunformity. One of
the generic drug product
ity. We maintained that an isoform variation in the active ingredient of
did not preclude a finding of active ingredient "sameness" for purposes of ANDA approval. We
noted in documents cited by the court that "complete chemical identification of all the carbohydrate variants in a protein product often is not possible or feasible.,,94 We stated "(i)ndeed, it
the same
usually is not even possible to 'assure by chemical analysis that different batches' of
product 'are identical at the level of the carbohydrate side chains' - including different batches
of Pergonal itself. ,,95
The D.C. Circuit upheld as reasonable the Agency's interpretation ofthe "sameness" statutory
requirement, as well as the Agency's interpretation ofthe word "identical" in 21 CFR
314.92(a)(I).96 The court concluded that the statute does not unambiguously require the term
"same as" to be defined as "complete chemical identity," noting that the statute says nothing at
demonstrate "sameness" nor about
all about the type of information an applicant must submit to
the type of information upon which the FDA may rely.97 The court characterized the sameness
provision as a "broad grant of discretion" to the Agency with respect to the information it mai
consider and noted that the phrase "must be read in the context ofthe kind of drug at issue.,,9
is based on
current scientific information and our knowledge of enoxaparin. Lovenox's enoxaparin is
naturally sourced and has batch-to-batch variability. As discussed at length, we have determined, based on relevant scientific data and information and our scientific experience and
expertise, that demonstration of active ingredient sameness for enoxaparin does not call for, nor
does the Act require, the ANDA applicant to establish sameness by meeting the requests set forth
in your petition regarding complete characterization of enoxaparin, manufacturing processes, or
clinical trials. We have concluded that if an ANDA applicant meets the five criteria (or stanthis response, this evidence demonstrates that the
dards for identity) described in section III of
the
generic
drug
product's
enoxaparin and Lovenox's enoxaparin wil be
molecular diversity of
equivalent and, therefore, provides sufficient information to conclude that the generic drg
Our decision here, as in Serono, takes into account the "kind of drug at issue" and
product's enoxaparin is the same as Lovenox' s enoxaparin.
94158 F.3d at 1318.

95Id.
96 Id. at 1321.
97 Id. at 1319.
98 Id. You also acknowledge that "the Act does not define what 'information' must be shown to establish sameness.
the active ingredient." (Aventis
Just what 'information' an ANDA must contain, therefore, depends on the nature of
Comments dated October 13,2004, p. 26.)
26
VI. SPECIFIC RESPONSES AND COMMENTS ON PETITION, SUPPLEMENTS,

AND RELATED COMMENTS
We have explained at length the basis for our decision on enoxaparin sameness. We address
below the arguments you raise in the Petition, the Supplements, and related comments.99
You assert that to demonstrate active ingredient sameness, the ANDA applicant must (1)
completely characterize all the different polysaccharides of enoxaparin by isolating, purifying,
and sequencing each of its unique polysaccharide chains and determining their relative abundance, (2) use Aventis's or an Aventis-equivalent manufacturing process, or (3) conduct clinical
trals to demonstrate equivalent safety and effectiveness. As summarized below, we have
thoroughly considered your arguments, and we reject them.
A. To Demonstrate Active Ingredient Sameness, ANDA Applicants for Enoxaparin Do
Not Need to Completely Characterize All the Different Polysaccharide Sequences

You claim that the only method by which an ANDA applicant could ensure its product would
have the same biological and clinical effects as enoxaparin (other than use of an A ventis-
equivalent manufacturing process or clinical trials to establish safety and effectiveness) is to

the different polysaccharide sequences and their relative amounts
characterize and compare all of
the polysaccharide chains
(Supplement No.3 at 2-3). You state that approximately 30 percent of
comprising enoxaparin have yet to be directly analyzed (without the benefit of complete
the limitations on current analytical technology
the sample) because of
enzymatic digestion of
polysaccharides
is difficult, laborious, and time(Petition at 3). You note that sequencing of
consuming, and you comment on the need to assure purity of samples (Supplement No. 3at 911 ). You state that A ventis has not been able to isolate those polysaccharide chain fragments of
molecular weight above 3,600 Da and that this portion remains unexplored by direct analysis
(Viskov Declaration, at 4). 100 You state that as technology continues to improve, investigation of
the unexplored portions of enoxaparin may yield additional unique and process-dependent
structural modifications with pharacological activity (Petition at 3). You also state that a
"generic product must contain those modifications (known or undiscovered) to be considered the
'same' as enoxaparin" (Petition at 3). You state that an ANDA applicant cannot claim to have
the same pharacological activity as enoxaparin simply because it has the same molecular
weight, anti-Xa activity, and/or anti-Xa/anti-lla ratio (Petition at 3-4, 20).
99 You take issue with the analytical techniques and data submitted by an ANA applicant in its May 13, 2004,
letter to Lester M. Crawford, D.V.M., Ph.D., then Acting Commissioner, FDA, that was subsequently forwarded to
the docket by cover letter dated June 1,2004 (Aventis Comments dated October 13,2004, and March 17,2005, and
Supplement No.2). FDA, as the Agency charged with reviewing ANAs, considers the data and information
submitted in ANAs before reaching any approval decisions. Accordingly, it is unnecessary to respond to your
assertions and judgments about the data and information submitted by any particular ANA applicant to the citizen
petition docket in this regard. Further, we are not generally permitted to disclose from ANAs confidential
commercial or trade secret information.
100 We note that Aventis has not sought to fully characterize enoxaparin even for those chains for which the
technology exists (Viskov Declaration at 4, noting "This does not imply that Aventis has fully characterized all of
those chains below 3,600 Da even though the technology exists to do so").
27
Based on current relevant scientific data and information and our scientific experience and
necessary to completely characterize all the different
expertise, we conclude that it is not
polysaccharide sequences of enoxaparin. Our finding of sameness for enoxaparin is not based
solely on the equivalence in molecular weight, anti-Xa activity, and/or anti-Xa/anti-lla ratio. We
conclude that the five criteria described in section III provide suffcient information to demonthe
strate active ingredient sameness. Such a showing demonstrates that molecular diversity of
generic drug product's enoxaparin and Lovenox's enoxaparn wil be equivalent and, therefore,
provides suffcient information to conclude that the generic drug product's enoxaparin is the
same as Lovenox's enoxaparin. As discussed at length in sections II, iv, and V, this conclusion
is supported by relevant scientific evidence, Agency precedent (e.g., heparin and hetastarch), and
law.
B. ANDA Applicants for Enoxaparin Do Not Need to Demonstrate That They Use the
Same Manufacturing Process as A ventis
We first address below your general claim regarding the need for using Aventis's manufacturing
process or the equivalent, and then we tum to your specific assertions and the information you
submitted.
1. Your general claim
You state that until enoxaparin has been fully characterized, we should refrain from approving
any ANDAs citing Lovenox as the RLD unless, among other things, the manufacturing process
used to create the generic drug product is deemed to be equivalent to A ventis' s manufacturing
process for enoxaparin (Petition at 1,10). You state that Aventis uses a processof-elimination
of uronic benzylic esters to manufacture enoxaparin and that the process creates a distinct drug
product with a unique chemical structure that is sensitive to specified temperature, base concenti.tion, and duration factors in the reaction (Petition at 10-11). In particular, you state that
different LMWHs manufactured through different depolymerization processes wil have

different ranges of oligosaccharde chains having a given molecular mass (Petition at 11 ). You
claim that Aventis's process results in particular saccharide sequences, which include distribution of specific structures as well as the type and arrangement of sacchardes within a given chain
(Petition at 11). You also assert that the manufacturing process creates specific processdependent structural "modifications (e. g., fingerprints) to enoxaparin' s chemical structure
(Petition at 11). You claim that use of a manufacturing process that differs from that for
Lovenox would likely result in a drug with a different oligosaccharide permutation, different
pharmacokinetics, and potentially dissimilar clinical activities. In particular, you claim that
differences in oligosaccharide length and sequence permutations would likely lead to varyng
absorption and elimination and different circulating anti-Xa/anti-lla ratios (Petition at
rates of
13_14).101 You maintain that until enoxaparin becomes fully characterized, the only way to
ensure that generic enoxaparin contains all the therapeutically significant structural features of
101 In addition to our response to your arguments in the text of this response, we note that any such differences that
you claim may affect the rates of absorption and elimination or different circulating anti-Xa/anti-lla ratios are
fuher ruled out by the active ingredient sameness criterion on equivalence of in vivo pharmacodynamic profies, as
explained in section III.E of this response.
28
enoxaparin (both known and yet to be discovered) is to duplicate Aventis's manufacturing

process or use an equivalent process (Petition at 20-21).
As discussed at length above, the Act and implementing regulations require the ANDA applicant
as a condition of approval to provide sufficient information to demonstrate that the active
the RLD. We conclude that the five
ingredient in the generic drug product is the same as that of
criteria described in section III of this response provide sufficient information to establish active
ingredient sameness for enoxaparin. One of the five criteria involves using the equivalent
heparin source material (i.e., heparin that is derived from porcine intestinal mucosa and that
meets USP monograph standards for Heparin Sodium USP) and the equivalent mode of depolythe benzyl ester derivative of
merzation (i.e., cleavage by alkaline -e1imination of
heparin) as
that used for Lovenox's enoxaparin.102 In addition to meeting these criteria, the ANDA applicant
the depolywould likely have to make appropriate adjustments to the operating conditions of
merization process to meet the other criteria described in section III. An ANDA applicant would
not need to know Aventis's exact manufacturing process parameters and conditions (e.g.,
depolymerization time, pH, and temperature) to manufacture the same active ingredient as
Lovenox's enoxaparin.103 There is no requirement that an ANDA applicant achieve active
ingredient "sameness" for enoxaparn by using the same manufacturing process as that used for
the RLD. You have not provided any information to show that an ANDA applicant's manufacproducing the same active ingredient as Lovenox's
turing process would be incapable of
enoxaparin within the meaningofthe Act and regulations.
Although you ask in your petition that ANDAapplicants for enoxaparin use an Aventisequivalent manufacturing process, it is unclear to what processes you are referrng. 104 The Act
and implementing regulations require an ANDA applicant, as a condition of approval, to ensure

that the methods used in, or the facilities and controls used for, the manufacture, processing, and
the drug are adequate to assure and preserve the identity, strength, quality, and purity
packing of
the drug.105 The two requirements - that an ANDA applicant demonstrate active ingredient
of
102 Lovenox's enoxaparin is derived by benzyl esterification of
the free carboxylate (uronic acid residue) of
hepari,
heparin is treated under alkaline
which is obtained from porcine intestinal mucosa. This benzylated ester of
conditions to induce -elimination, which results in polysaccharide chain cleavage. Likewise, we expect the ANA
applicant to use hepari derived from porcine intestinal mucosa and to depolymerize this source material though
cleavage by alkaline (chemical) -elimination of
hepari.
103 As a general matter, manufacturing information is considered confidential commercial and/or trade secret
information, which cannot be disclosed by the Agency.
the Lovenox manufacturing

process have remained unchanged (Petition at 11). On the other hand, you state that between March 1996 and April
which were drug product related and the remaining eight of
2004, you submitted 24 CMC supplements, sixteen of
which were not solely drug product related (Aventis October 13,2004, Response to Comment at 20-22).
105 See section 505(j)(4)(A) of
the Act, and 21 CPR 314.l27(a)(1) and 3l4.94(a)(9)(i) (referencing 314.50(d)(l)).
The ANA applicant submits the same tye of information on chemistr, manufacturing, and controls as NDA
applicants, including the following: "A full description of the drug substance including its physical and chemical
characteristics and stability; the name and address of its manufacturer; the method of synthesis (or isolation) and
purification of the drug substance; the process controls used during manufacture and packaging; and the specificathe drug substance and the bioavailability of
tions necessary to ensure the identity, strength, quality, and purity of
the drug products made from the substance, including, for example, tests, analytical procedures, and acceptance
104 You note that since the initial development of enoxaparin in 1981, the steps of
criteria relating to stability, sterility, paricle size, and crystallne form. The application may provide additionally for
the use ofaltematives to meet any of
these requirements, including alternative sources, process controls, and
29
sameness. and submit designated information on the manufacturing process - are independent
statutory and regulatory requirements and are addressed in two different sections of the statute
an ANDA applicant use the
and regulations. Neither the Act nor FDA regulations require that
same methods used in, or the facilities and controls used for, the manufacture, processing, and
the
of
the RLD to assure and preserve the identity, strength, quality, and purity
packing of
106 As discussed elsewhere, once we conclude that an ANDA for enoxaparn meets
generic drug.
the requirements for ANDA approval, the generic enoxaparin and Lovenox can be substituted
with the full expectation that generic enoxaparn wil produce the same clinical effect and safety
profie as Lovenox.
2. Your specifc assertions
a. The 1,6 anhydro ring structure
You specifically request that we not approve generic enoxaparin unless the generic product
its
between 15 to 25 percent of
contains a 1,6 anhydro ring structure at the reducing ends of
oligosaccharide chains (Petition at 1). You state that Aventis's manufacturing process results in
formation of the 1,6 anhydro ring during the -e1imination depolymerization process, and the
the 1,6 anhydro ring is sensitive to Aventis's process conditions
formation or frequency of
(Petition at 13). You state that unlike enoxaparin's other strctural fingerprints, you have
conducted preclinical tests on the 1,6 anhydro ring structure which lead you to conclude that this
structure has pharacological activity at the 15 to 25 percent frequency and might have an effect
on inflammation, smooth muscle cell proliferation, angiogenesis (potentiation of acidic fibroblast

growth factor (aFGF)-induced endothelial cell proliferation), coagulation and thrombosis,
pharmacokinetics, and safety profile (Petition at 14-19). You state that A ventis' s scientists
constructed two LMWHs similar to enoxaparin in molecular weight, anti-Xa activity, and antiXa/anti-lla ratio, but with dissimilar 1,6 anhydro ring content (i.e., .:7 percent 1,6 anydro
LMWH and 40 to 50 percent 1,6 anydro LMWH) (Petition at 14). You state that Aventis
compared these two alternative LMWHs with enoxaparin to assess various aspects of pharmacological activity and gauge the pharmacological relevance of the 1,6 anhydro ring structure
(Petition at 15).
As discussed in section III, if an ANDA applicant for enoxaparin meets the five criteria, this
the generic drug product's enoxarobust showing demonstrates that the molecular diversity of
parin and the Lovenox's enoxaparin wil be equivalent, including with respect to the 1,6 anhydro
ring structure. 107 This wil also result in meeting the standards for identity described in the USP
monograph for enoxaparin sodium, which states that "(a )bout 20 percent of the materials contain
analytical procedures. Reference to the current edition of
the U.S. Pharmacopeia and the National Formulary may
satisfy relevant requirements in this paragraph."
106 We need not decide whether for a different active ingredient (i.e., an active ingredient other than enoxapari) the
same methods, facilities, and controls used for the manufacturing, processing, and packing as those used for the
the generic drug; here, we
conclude that this is not the case for enoxapari.
107 Recognition of
these fingerprints, as you also acknowledge (Petition at 3), is possible due to recent advances in
the field of analytical technology (see also section III.e).
RLD would be needed to assure and preserve the identity, strength, quality, and purty of
30

a 1,6 anhydro derivative on the reducing end of the chain, the range being between 15 and 25
percent."io8
We also note that you have not provided adequate information for us to conclude that the 1,6
anhydro ring structure is clinically significant, as you claim. In considering your arguments, we
the articles and information submitted with your petition and supplements.
have reviewed all of
You refer to in vitro and preclinical studies to support your claims regarding the purported
contributions of the 1,6 anhydro ring structure at a frequency of 15 to 25 percent. Given the
quantum of articles and reports submitted, it is not possible to address each one in this response,
nor do we find it necessary. Nonetheless, we provide below some examples to convey the nature
of
the materials submitted.
Inflammation
the data you submitted on their face do not fully support the conclusions you attempt to
Some of
draw regarding the purported effect ofthe 1,6 anydro ring structure on inflammation. For
example, you refer to the Petitet report to support your claims regarding the purported contribu-
tions of the 1,6 anhydro ring structure to the anti-inflammatory activity of enoxaparin (Petition,
Appendix A). Although the report includes data on the inflammatory properties of enoxaparin,
.c7 percent 1,6 anhydro LMWH, 40 to 50 percent 1,6 anhydro LMWH, and other saccharide
fragments, the conclusion drawn by the author relates only to the anti-inflammatory properties of
the 1,6 anydro ring strcture to the anti-inflammatory
enoxaparin, and not to contributions of
activity of enoxa~arin (Petitet report at iv). Additionally, you cite another study that used CD54
expression to account for enoxaparn's anti-inflammatory effects, but enoxaparin's effect on
expression was found not to be dependent on the 1,6 anydro group content (Supplement
No.4 at 4).
CD54
I
i
Smooth muscle proliferation

Some of your statements on the purported effect of the 1,6 anhydro ring structure on smooth
muscle proliferation are merely speculative. For example, you state that "a product claiming to
be enoxaparin that lacked the 1,6 anhydro ring structure (or had it in a different concentration)
could have a different effect on inhibition of SMC (smooth muscle cell) proliferation" (Supplethe data you submitted on their face do
not fully support the conclusions you attempt to draw with respect to smooth muscle proliferation. For example, while the Diley & Little authors (Petition, Appendix A) reported that both
the.c 7 percent 1,6 anhydro LMWH and the 40 to 50 percent 1,6 anhydro LMWH exhibited
distinctive inhbitory properties of smooth muscle cell proliferation (Diley & Little at 7), the
same authors also indicated that variability was an issue in these experiments. These authors
indicated that in these experiments it was difficult to determine whether there was a consistent
ment No.1 at 10) (emphasis added). Further, some of
these compounds may
dose-response relationship and suggested that more extensive analysis of
provide more accurate data (Diley & Little at 9).
108 See USP 32-NF 27 monograph for enoxaparin sodium (offcial on December 1,2009).
31
Angiogenesis
Some of
the 1,6 anhydro ring structure on
your statements regarding the purported effect of
angiogenesis are also speculative. For example, you state that "a product that claimed to be
enoxaparin but lacked the 1,6 anydro ring structure (or had it in a different concentration than is
found in enoxaparin) could have a different effect on stimulation of angiogenesis than does
the data you submitenoxaparin" (Supplement No.1 at 10) (emphasis added). Further, some of
ted on their face do not fully support the conclusions you attempt to draw with respect to
angiogenesis. For example, while the report from von Specht (Petition, Appendix A) indicates
that the.: 7 percent 1,6 anhydro LMWH is significantly more active than the 40 to 50 percent 1,6
line (baby hamster kidney cells) (von Specht at
anhydro LMWH when assayed through BHK cell
10), the same author indicates that because of the high standard deviations obtained in cell
culture assays, they were unable to calculate a rank order of the LMWHs tested. Moreover, the
same author's test results (using the chorion allantois membrane (CAM) assay (at 8 iig of
L WMH dose) to assess angiogenesis) suggest that the 40 to 50 percent .1,6 anhydro LMWH (as
opposed to the.: 7 percent 1,6 anhydro LMWH) was more active (von Specht at 10), which
appears to contradict the results obtained from the BHK cell test.
Coagulation and thrombosis
Again, some of your statements regarding the effect of the 1,6 anhydro ring strcture and
bleeding profiles are merely speculative. For example, you state that the "presence of
the 1,6
anhydro ring structure appears to change the anti-coagulant potency and antithrombic effect of
enoxaparin, which may. . . lead to different bleeding profiles in patients" (Supplement No. 1 at
8) (emphasis added). In addition, some of
the data you submitted are on their face inconsistent
with the conclusions you attempt to draw with respect to coagulation and thrombosis. For
example, you concede that your tests did not show a statistically significant difference between
enoxaparin (containing the 1,6 anhydro ring at 15 to 25 percent frequency) and the two alternathe 1,6anhydro ring structure) with respect to
tive LMWHs (containing other frequencies of
activated partial thromboplastin time, prothrombin time, thrombin generation, anti-Xa activity,
another study that examined the
anti-I1a activity, or thromboelastography (Petition at 19). In
capacity of enoxaparin and its components to attenuate tissue factor (TF) expression and
subsequent activation of coagulation on the cell surface, you found that the 1,6 anydro group
was not an important determinant of enoxaparin's activity in this assay (Supplement NO.4 at 7).
You also claim that a recent study demonstrates that TFPI inhibition reversal is another processdependent biological property of enoxaparin that may have clinical significance (Supplement No.
2 at 3-4). Specifically, you state that the study demonstrates that both polysaccharide chain
length and 1,6 anhydro ring structure concentration affect enoxaparin's reversal ofTFPI
inhibition (Supplement No.2 at 4). You state that TFPI release plays a major role in neutralizathe TF/factor VIla complex initiating coronary thrombosis after artery injury or plaque
tion of
rupture (Supplement NO.2 at 2-5). Therefore, you state that if an ANDA applicant does not use
an A ventis-equivalent manufacturing process, the enoxaparin "might" exhibit a different antithrombotic profie than enoxaparin (Supplement NO.2 at 5). We note that your conclusions are
not based on relevant data, but instead are hypothetical outcomes. As the authors noted,
32
"(fJurther studies are needed to define the role and mechanisms of 1,6anhydro in endothelial
TFPI under varous conditions" (Supplement No.2, Exhibit A).
Pharmacokinetics
Your claim regarding the purported contributions of the 1,6 anhydro ring structure at a frequency
of 15 to 25 percent with respect to enoxaparin pharmacokinetics (in vivo pharmacodynamic
profiles) is based upon studies in beagle dogs, as opposed to studies in humans. We further note
that even these pharmacokinetic (in vivo pharmacodynamic profies) data on their face do not
support the conclusions you attempt to draw. For example, you state that -:7 percent 1,6 anhydro
LMWH showed a statistically significant increase in plasma anti-Xa activity and anti-lla activity
compared to enoxaparin (Petition at 18, Appendix A; Report DMPKIFRA2003-0029 (DMPK
Report), pp. 20-25). In your report, you state that for the -:7 percent 1,6 anhydro LMWH after
one subcutaneous administration in dogs, there were statistically significant differences in certain
parameters compared to enoxaparin (DMPK Report, pp. 20). However, you also state that other
anti-Xa and anti-lla pharacokinetic parameters were not statistically significant. Further, you
the 40 to 50 percent 1,6 anydro ring LMWH, no statistically
state that after administration of
significant differences between the pharmacokinetics of that compound and enoxaparin were
observed (DMPK Report at 25).
Safety profie
Some of the data you submitted are on their face inconsistent with the conclusions you attempt to
draw with respect to safety profie. For example, heparin-induced thrombocytopenia (HIT) is a
heparin therapy; however your in vitro studies showed that the
potentially fatal complication of
1,6 anhydro ring structure does not modify enoxaparin's HIT cross-reactivity (Supplement No.1
at 11). We also note that your statements on the purported effect of the 1,6 anydro ring
structure and safety profile are merely speculative. For example, you state that "a generic
hemorrhagic
product that ... did not contain the 1,6 anhydro ring may pose an increased threat of
complications in patients" (Petition at 19) (emphasis added).
Although you speculate that the studies submitted "likely bear clinical significance" (Petition at
3), you have not provided any clinical studies to support your claim. Accordingly, you have not
provided sufficient evidence upon which we can reasonably conclude that the 1,6 anhydro ring
has the pharmacological activity or clinical signifistructure at the 15 to 25 percent frequency
109
cance in humans that you claim.
. 109 While you submitted information on the 1,6 anhdyro strcture to support labeling changes for Lovenox (NA
20-164/S-055 at 3), the review division concluded that additional studies would be needed to "establish the clinical
importance of
this chemical characteristic." See Kathie M. Robie-Suh, M.D., Ph.D., Medical Team Leader, Review
20-164/S-055 dated July 22,2004.
pdf)
(htt://www.accessdata.fda.gov/drugsatfda _ docs/ndaI2004/020 164_ S055 _ Lovenox _Approval_ Package.
33
b. AT-III binding oligosacchardes 110
You assert that LMWHs inhibit coagulation by binding to AT -III; a plasma protein synthesized
in the liver and other cells (Supplement NO.1 at 3). You state that LMWH interaction with ATIII is mediated by a specific pentasaccharide sequence that is distributed across 15 to 25 percent
of LMWH polysaccharide chains (Supplement No. 1 at 3). You state that you discovered that
process-dependent varations in saccharide sequence within a given oligosaccharide affect the
oligosaccharide's affinity for AT-III (Supplement No.1 at 4). You state that the differences in
. the disaccharide sequences occur depending upon where the unfractionated heparin is cleaved
during the manufacturing process, and A ventis' s process results in three main process-dependent
Aventis conducted in vitro
octasaccharde sequences (Supplement NO.1 at 5). You state that
its three octasaccharide
affinity and the anti-Xa activity of
experiments to measure the AT-III
sequences (Supplement No. 1 at 5). You state that these findings show that different octasaccharides in enoxaparin do not have identical in vitro anti-Xa activity and there can be considerable
variation in affnity for AT -III (Supplement No.1 at 6-7). You maintain that a recent study
shows that these process-dependent AT-III binding sites are found not only in enoxaparin's
octasaccharide fractions, but in all ofthe drug's oligosaccharide fractions (Supplement NO.2 at
the anti-Xa
7-8). You claim that sequence variations might cause differences in the half-lives of
of
bleeding safety and antithrombotic effectiveness
activity, leading to different profiles
(Supplement No. 1 at 4-7).
what you refer to as the "classical" AT-III binding

sequence in enoxaparin is influenced by the nature of the particular saccharide units flanking that
sequence, and that the use of a surrogate for the classical AT-III binding sequence such as the
one you reference cannot adequately predict AT-III binding activity (Supplement NO.3 at 36).1 I ! You also claim that some polysaccharids have AT-III binding affinity even though they
lack the classical AT-III sequence (Supplement NO.3 at 7-9). You state that any differences
could influence antithrombotic activity (Supplement NO.3 at 8). You state that these apparent
You state that the AT-III binding affnity of
sacchardes in enoxaparin (Supplement
findings underscore the need to characterize all the oligo
NO.3 at 6 and 7). You also discuss Dr. Boudier's findings that non-ionic interactions are
important in the binding of octasaccharide D with AT-III. You indicate this demonstrates that
"small changes in sugar structures flaning the classical AT-III binding sequence can have large
effects on AT-II binding" (Supplement No.4 at 10).
You cite in vitro studies to support your claims regarding the possible contributions of what you
your
refer to as process-dependent sequences flanking the AT-III binding sites. Many of
saccharides are merely speculative.
statements regarding process-dependent AT-III binding oligo
For example, you referto a recent study that you claim demonstrates that enoxaparin has a
process-dependent inhibitory effect on factor VIla generation, which you state is linked to
arterial thrombogenesis (Supplement NO.2 at 5-7). You state that the study demonstrates that
110 The terms "process-dependent AT-III binding oligosaccharides" and "process-dependent sequences flankg the
this response.
i i i You state that researchers collaborating with one ANA applicant have published this method for detecting this
AT-II binding oligosaccharides" are used interchangeably for puroses of
purported surogate for AT-II binding activity (Supplement No.3 at 6). We note that we would not consider a
generic drug product's enoxaparin to be the same as Lovenox's enoxaparn based solely upon the surrogate you
describe. As discussed at length, we consider the generic drug product's enoxaparin to be the same as Lovenox' s
enoxapari if the five criteria described in section II are met.
34
the concentration of AT-III binding sites within enoxaparin influences factor VIla generation
factor VIla
inhibition (Supplement NO.2 at 6). You state that enoxaparin's inhibition of
generation represents another biological property of enoxaparin with "possible" clinical significance (Supplement No.2 at 6). You speculate that a generic enoxaparin that did not use an
equivalent manufacturing process "might" contain a different distribution and structue of AT-III
binding sites, and this in tum "could" lead to a different effect on inhibition of factor VIla, and
such a product "might" exhibit a different effect on arterial thrombosis which "could" have
clinical consequences (Supplement No.2 at 5-7). Even the study authors propose four areas of
future study that would be needed for a "complete description and comparison of the oligosaccharides effect on the blood coagulation process" (Supplement No.2, Appendix B). Further, Dr.
Boudier in his studies of octasaccharide D acknowledges that both the small size of these
confer a substantial specificity (especially
products and their high affinity for AT-II "may"
product D), a key advantage in pharmacology (Supplement No.4, Appendix F).
We conclude that you have not provided suffcient information upon which we can conclude that
saccharides are clinically significant.
what you refer to as process-dependent AT-III binding oligo
Although you speculate as to the clinical significance of your studies, you have not provided any
clinical studies to support your claim. Without data from clinical studies, we cannot reasonably
conclude that what you refer to as the process-dependent AT-III bindingoligosaccharides and
"nonclassical" AT-III binding sequences you reference are pharmacologically and clinically
active in humans.
Based on the available scientific evidence, we conclude that in conjunction with showing
conformance to the other criteria described in section III, performing a sequence analysis on a
subset of oligosaccharides in enoxaparin (having sequences that are sensitive to varations of
process conditions used during depolymerization) and demonstrating that this subset of oligosacsaccharides in Lovenox's
charides has sequence(s) equivalent to a comparable subset of oligo
the generic drug prodenoxaparin provides sufficient evidence that the molecular diversity of
uct's enoxaparin and Lovenox's enoxaparin wil be equivalent, including the process-dependent
AT-III binding oligosaccharides and "nonclassical" AT-III binding sequences that you describe.112 Equivalent molecular diversity demonstrates sameness for enoxaparin.
c. Other structural fingerprints
In your petition, you state that you have identified several other structural fingerprints in
enoxaparin that you claim "may contribute to its pharmacological activity" (Petition at 12). You
state that these include oligosaccharides with odd-numbered saccharide units, galacturonic acid
moieties, and epimerized reducing ends in a mixture of 70 percent glucosamine and 30 percent
mannosamine (Petition at 12). You concede that Aventis "has not yet assessed the pharmacological activity ofthe(~eJ other fingerprints" (Petition at 13). You nonetheless speculate that
112 As discussed in section III, the ANA applicant can ensure that the depolymerization process conditions used in
the generic drug product's enoxaparin have a chemical selectivity of cleavage that is equivalent
the depolymerizationprocess used to manufacture Lovenox's enoxaparin. Thus, the
ANDA applicant can ensure that the site(s) where the unfractionated heparin is cleaved wil be equivalent between
the generic drug product's enoxaparin and Lovenox's enoxpari. We can expect the generic drug product's
enoxaparin to be equivalent to Lovenox's enoxaparin with respect to what you refer to asprocess-dependent AT-II
binding oligosaccharides.
the manufacture of
to the chemical selectivity of
35
these fingerprints. . . represent significant structural modifications and wil
"(mJany of
likely
prove to be pharmacologically and clinically active" (Petition at 13).

We have concluded that you have not submitted any data to show that these other structural
modifications are pharacologically or clinically active. Therefore, your statements are
conclusory and speculative and do not support your claims. As stated above, if an ANDA
applicant meets the five criteria described in section III of this response, this robust showing
demonstrates that the molecular diversity ofthe generic drug product's enoxaparin and
Lovenox's enoxaparin wil be equivalent - including with respect to the odd-numbered
saccharide units, galacturonic acid moieties, and epimerized reducing ends having a mixture of
glucosamine and mannosamine structures. Equivalent molecular diversity demonstrates
sameness for enoxaparin.
C. ANDA Applicants Are Not Required to Submit Clinical Trials to Establish the
Safety and Effectiveness of Enoxaparin
Absent full characterization or use of Aventis's or an equivalent manufacturing process for
enoxaparin, you state that we cannot consider the generic drug product to have the "same" active
ingredient as enoxaparin and, therefore, we must require a demonstration of equivalent safety
and effectiveness through clinical testing. In such cases, you state that section
the Act prohibits FDA from approving the ANDA unless the applicant
505G)(2)(A)(ii)(II) of
establishes the safety and effectiveness of its enoxaparn product through clinical trials (Petition
at 1,21).11
the Act provides that an ANDA

applicant must submit "information to show that the active ingredient of the new drug is the
the listed drug.,,1l4 The Act also provides that we must approve an ANDA
same as that of
Your claim is without merit. Section 505(j)(2)(A)(ii)(I) of
ingredient
IS Section 505(j)(2)(A)(ii) of the Act does not, as you claim, prohibit us from
unless, among other things, it includes "insufficient" information to show active

i
sameness.
approving an ANDA that does not include clinical trials to demonstrate the safety and effectiveness of the enoxaparin product. This section of the Act does not include any language specifying
the type of information needed to demonstrate active ingredient sameness, and we find clinical
studies to demonstrate safety and effectiveness are not necessary here. The requirement you
propose would be contrary to one of the principal purposes of the ANDA statutory approval
bringing
provisions, which is "to encourage competition by decreasing the time and expense of
i 13 You also assert that enoxaparin is similar to a biologic (Petition at 8). You question whether the generic drug
the
approval model is appropriate (Petition at 8). Lovenox is not a biological product licensed under section 351 of
the
Act.
Although
it
is
not
a
Public Health Service Act (PHS); rather, Lovenox is approved under section 505 of
foregone conclusion that a showing of active ingredient sameness can be made for all drgs approved under section
the Act, we conclude that active ingredient sameness can be demonstrated for enoxaparin. FDA can address
505 of
your arguments regarding the specific characteristics of enoxapari without wholesale rejection of the statutory
the Act. As with certain other naturally sourced
process applicable to ANA applications under section 505 of
the Act is
products, like heparin and hetastarch, approval of ANAs for enoxaparin under section 505) of
appropriate.
i 14 You specifically refer to section 505(j)(2)(A)(ii)(II) of the Act, which pertains to drgs containing more than one
the Act, which pertains to

single active ingredient drugs. Enoxaparin sodium injection is a single active ingredient drg.
115 See section 505(j)(4)(C)(i) of
the Act.
active ingredient. Elsewhere in your petition you refer to section 505(j)(2)(A)(ii)(I) of
36

generic drugs to market, and thereby to provide the public with low cost drugs."! 16 Further, the
conduct of duplicative studies raises ethical concerns because it could subject humans and
animals to medically or scientifically unjustified testing.
As discussed and as you acknowledge (Aventis October 13, 2004, Comments at 26), we have
considerable discretion to determine what type of information is sufficient to demonstrate active
ingredient sameness for ANDA approval. Based on our scientific experence and expertise and
relevant scientific information, we have determined that ANDA applicants can provide sufficient
information to demonstrate the sameness of enoxaparin by showing that the generic drug
product's enoxaparin meets the five criteria (or standards for identity) discussed in section III of
this response. Submission of ANDAs under section 5050) ofthe Act is an appropriate pathway
for approval of generic enoxaparin.
D. Contrary to Your Assertions, Approval of an ANDA for Enoxaparin Is Consistent
With FDA Precedent

1. Pergonal
You state that the case of enoxaparn is distinguishable from previous cases in which we have
concluded that a proposed generic drug was the same as the RLD despite differences in active
ingredient chemical structure (Petition at 21). Specifically, you state that we approved an ANDA
for a generic version ofPergonal (with active ingredients FSH and LH), even though the generic
version had a different isoform of FSH than Pergonal (Petition at 21 ). You state that we
concluded the isoform variation was not clinically significant for the product's int~nded uses
and, therefore, it did not preclude a sameness finding (Petition at 21 ). You recognize that in
Serono, the Court of Appeals held that we are entitled to deference in our interpretation of the
"same as" in the statute (Petition at 21). However, you claim that, unlike the isoform
variation at issue in Serono, preclinical tests show that the 1,6 anhydro ring structure at a
frequency of 15 to 25 percent in enoxaparin "may well" have clinical significance (Petition at
meaning of
21). You also state that, unlike Pergonals active ingredient at the time of ANDA submission,
Lovenox is not fully characterized (Petition at 22). Therefore, you claim that we
cannot ensure
that a generic drug product's enoxaparin is the same as Lovenox' s enoxaparin unless the product
is manufactured using a process equivalent to the Lovenox process (Petition at 22).
the approval ofPergonal does not support your arguments with respect to
Lovenox. Consistent with Serono, our finding of active ingredient sameness is based on relevant
scientific information and is specific to each active ingredient. Although enoxaparin and
menotropins (FSH and LH) are heterogeneous mixtures of molecular entities, the active ingredients have different origins and are composed of entirely different molecular structures. Enoxa-
Your discussion of
parn is a heterogeneous mixture of oligosaccharides produced through alkaline depolymerization of
the benzyl ester of
menotropins are amixture of
heparin derived from porcine intestinal mucosa, whereas Pergonal's

postprotein isoforms ofFSH and LH derived from the urine of
menopausal women.
ii6 See 54 FR 28872 at 28874 (July 10, 1989).
37
For Pergonal, we expected the active ingredient in the generic version to have the same primary
tobatchsource material), potency, and degree of
structure (assured by using the same natural
batch uniformity as the innovator's active ingredient. We concluded that any differences in
isoform variation between the generic drug product's active ingredient and Pergonals active
ingredient w,ere not clinically significant. For enoxaparin, we have concluded that the five
the USP monograph for enoxacriteria described in section III (which result in satisfaction of
parin sodium, as well as additional standards that are material to enoxaparn's sameness) are
suffcient to demonstrate enoxaparin sameness. Although you have not provided adequate data
to demonstrate that the 1,6 anhydro ring structure or other strctures have pharmacological
or clinical significance in humans as discussed in section VI, our five criteria approach
the generic drug product's
for enoxaparn sameness demonstrates that the molecular diversity of
enoxaparin and Lovenox's enoxaparn wil be equivalent, including with respect to the 1,6
activity
anhydro ring structure. Equivalent molecular diversity provides sufficient information to
conclude that the generic drug product's enoxaparin is the same as Lovenox's enoxaparin.
the molecular entities
that comprise enoxaparin and menotropins (FSH and LH)), it is reasonable and appropriate for
the Agency to take into account their respective molecular diversity and form conclusions and
approaches specific to each active ingredient.
Based on relevant scientific evidence (and the vastly differing structures of
2. Premarin
of enoxaparin is highly analogous to that of Premarin, a conjugated
estrogen product for which the innovator claimed the active ingredient had not been adequately
characterized and, therefore, could not be duplicated by ANDA applicants (Petition at 22). You
note that in 1997, FDA concluded that Premarin must be characterized before synthetic generic
Premarins could be approved (Petition at 23). You also acknowledge that FDA stated that it
would approve a generic version of Premarin that was made using the same source material
Pre
marin (Petition at 24). You state that unlike conjugated
(pregnant mare's urine) as that of
estrogens, the composition of enoxaparin is not solely a factor ofthe source material used, but is
You state that the case
allege that for
also highly dependent on the manufacturing process (Petition at 23). You
enoxaparin, the process makes the product (Petition at 24). You also claim that the discovery of
the 1,6 anhydro ring to enoxaparn's activity is analogous
the pharacological contributions of
to the discovery of
the contributions of A (8,9) dehydroestrone sulfate (DHES) in Premarin
(Petition at 23). You state that the 1,6 anhydro ring in enoxaparin shows that Aventis's manufacturing process creates biologically relevant structures that do not exist in the natural source
material (Petition at 24). You maintain that Aventis ensures that all significant structural
modifications are consistently present in Lovenox through use of its tightly controlled manufac-
turing process (Petition at 24).
The parallels you attempt to draw between enoxaparin and Premarin are misplaced. Our finding
of active ingredient sameness is based on relevant scientific information and is specific to each
active ingredient. Although both enoxaparin and conjugated estrogens are heterogeneous
mixtures of molecular entities, these products are derived from different origins and are commixture of
posed of entirely different molecular structures. Enoxaparin is a heterogeneous

the benzyl ester of
oligosaccharides produced through alkaline depolymerization of
38
heparin
derived from porcine intestinal mucosa, whereas Premarin's conjugated estrogens are a mixture
of steroids derived from pregnant mare's urine.
For Premarin, we determined that we could not make a finding of active ingredient sameness for
marin because the Premarin active ingredient had not been adequately
Pre
characterized.II7 As you also acknowledge (Petition at 23), our conclusion for Premarin did not
preclude a finding of sameness for a generic drug product's active ingredient if the active
ingredient was derived from the same natural source material as the RLD.II8 Our conclusions
regarding active ingredient sameness for both synthetic and naturally derived Premarin were
~ased on relevant scientific information available at the time and our knowledge of the active
ingredient.
synthetic versions of
For enoxaparin, we have concluded based on relevant scientific evidence that there is sufficient
information to assess whether generic enoxaparin contains the same active ingredient as
Lovenox. Specifically, we conclude that the five criteria described in section III are sufficient to
conclude enoxaparin sameness. As described in section III, these criteria - including the use of
equivalent heparin source material - are designed to take into account the molecular diversity of
enoxaparn. As described in sections II and VI.B.2.a, we can expect that the generic drug
product's enoxaparn would contain the 1,6 anydro ring structure at a frequency of 15 to 25
percent. 119 Based on the available relevant scientific evidence (and the differing structures of the
molecular entities that comprise enoxaparin and conjugated estrogens), it is reasonable and
for the Agency to take into account their respective molecular diversity and form
appropriate
conclusions and approaches specific to each active ingredient.
3. Hyaluronidase
You state that our comments in response to a citizen petition submitted by 1ST A Pharmaceuti-
cals, Inc. (1ST A) concerning marketing exclusivity for its naturally sourced hyaluronidase
product, Vitrase, support denial of approval of generic enoxaparn (A ventis March 16, 2006,
petition, ISTA requested that we reverse our

decision to grant a 5-year period of marketing exclusivity for Vitrase and instead grant a 3-year
eomments at 3). You note that in the citizen
11762 FR 42562 at 42564-42572 (August 7,1997). We concluded at that time that there was insuffcient information to show that the active ingredients of the synthetic conjugated estrogen tablets were the same as the active
ingredients in the reference listed drug (derived frm natural source material- pregnant mare's urine). We could
not determine which constituents in the product were responsible for making clinically meaningful contributions to
the two most abundant estrogens,
its therapeutic effects. Among other things, we noted that (1) the contrbution of
Pre marin was not well
sodium equilin sulfate and sodium estrone sulfate, to the overall estrogenic potency of
understood; and (2) although the available evidence indicated that sodium L\ 8,9-dehydroestrone sulfate (DHES) was
active and contributed to the estrogenic potency, the clinical significance of
that contribution had not been
determined.
i 18 Memorandum to Douglas L. Sporn, Director, Office of Generic Drugs from Director, CDER, Subject:
Approvability ofa Synthetic Version of
Pre
marin, May 5,1997.
htt://ww.fda.gov/Drugs/DrugSafetvInformationbyDrugClass/ucm168836.htm.
the 1,6 anhydro ring structure in enoxaparin to the DHES in Premarin is
somewhat misplaced because, as stated above, you have not provided any clinical data on the potential significance
119 We also note that your comparison of
of
the 1,6 anyhdro ring strcture (whereas there were clincal data regarding DHES and its contribution to
Premarin).
39
Docket No.FDA-2003-P-0273
exclusivity period (A ventis March 16, 2006, Comments at 2). You also note that in a letter to
ISTA dated October 25,2005, we denied ISTA's request. 120
You maintain that in rejecting ISTA's request, we relied heavily on the fact that hyaluronidase is
not fully characterized (Aventis March 16, 2006, Comments at 2). You also note that our
response to ISTA'stated that because hyaluronidase was not fully characterized, we did not know
whether hyaluronidase products contained any previously approved active moieties. You further
state that because enoxaparn, like hyaluronidase, is not fully characterized, there is no way for
us to determine whether the active ingredient specified in an ANDA for enoxaparin is the same
as the active ingredient in Lovenox.121
Our hyaluronidase decision focused primarily on the appropriate application of certain exclusivity provisions (including the phrase "previously approved active moiety") to applications for
the Act. As you note, we stated in our
hyaluronidase submitted under section 505(b)(2) of
the Agency has insufficient information to
response to the ISTA petition that "(g)enerally, if
know whether a product contains a previously approved active moiety, the applicant would be
required to submit an NDA containing substantial clinical safety and effcacy data.,,122 We
concluded that we had "no information showing that any hyaluronidase products have been
adequately characterized to enable the Agency to determine whether they contain a previously
approved active moiety.,,123 We also referred to hyaluronidase as an unusual circumstance and
ultimately concluded that it was appropriate to grant 5 years of exclusivity to the applications at
issue. Although the hyaluronidase decision you reference focused primarily on the question of
exclusivity, we stated in a footnote that "FDA currently would also be unlikely to consider these
products (at issue) to have the same active ingredient for purposes of approving an application
under section 5050) ofthe ACt.,,124
hyaluronidase are
misplaced and not relevant. While both enoxaparin and hyaluronidase are heterogeneous
Nonetheless, the parallels you attempt to draw between enoxaparin and
mixtures of molecular entities, these two active ingredients are derived from different origins and
are composed of entirely different molecular structures. Enoxaparin consists of a heterogeneous

the benzyl ester of
heparin derived from porcine intestinal mucosa, whereas hyaluronidase is a mixture of glycosylated protein isoforms (isoenzymes) that is derived from either naturally sourced tissue - ovine
tissue (Vitrase) or bovine tissue (Amphadase, Hydase) - or through recombinant means (Hylenex).
mixtue of oligosaccharides produced through alkaline depolymerization of
120 Letter to Marvin 1. Garrett, Vice President, ISTA Pharmaceuticals, Inc., from Steven K. Galson, FDA, Docket
A). (The docket number for ths citizen petition was
25, 2005 (Response to 1ST
No. 2005P-0134/CP1, October
FDA's
transition
to its new docketing system.)
changed to FDA.2005-P-0005 as a result of
121 In a previous comment, you stated that "Aventis has never argued that FDA may not approve a generic
Enoxaparinproduct until Enoxapari is fully characterized." Aventis October 13, 2004, Comments at 28-29.
122 Response to ISTA at 9 (ISTA's petition concerned whether an active moiety had been previously approved for
purposes of determining marketing exclusivity under 21 CFR 314.108. for an application submitted under 505(b )(2)
of the Act, whereas your petition concerns the sameness of the active ingredient of a generic drug product under
the Act).
3l4.92(a)(1) for an ANA submitted under section 505(j) of
123 Response to ISTA at 5.
124 Response to 1ST A at 11.
40
For hyaluronidase, we noted that "the active ingredient in (hyaluronidase) has not yet been
sufficiently characterized to permit the Agency to conclude that another hyaluronidase product
the decision, we noted that we lacked
has an identical active ingredient.,,125 At the time of
sufficient information to conclude active ingredient sameness for hyaluronidase. In contrast,
enoxaparin has been adequately characterized for naturally sourced generic enoxaparin and there
is sufficient information for enoxaparin to conclude that the generic drug product's active
ingredient is the same as Lovenox's enoxaparin. Based on the available relevant scientific
the molecular entities that comprise enoxaparin and
evidence (and the differing structures of
hyaluronidase), it is reasonable and appropriate for us to take into account their respective
molecular diversity and form conclusions and approaches specific to each active ingredient.
E. Current Approval Requirements for ANDAs for Enoxaparin are Scientifcally
Appropriate and Address the Safety Proie of Enoxaparin.

The North American Thrombosis Foru (NATF), for example, states that current guidelines for
generic drug approval may not be applicable for the approval of therapeutic agents that are
biologically based. In particular, NATF and other comments state that there is the potential for
unanticipated adverse events or immune response, and that clinical testing is essential (February
Hospital Medicine
22,2010 and March 2,2010, NATF Comments; May 26,2010, Society of
Comment; and May 28, 2010, Victor Tapson, Duke University Medical Center Comment).
the generic product to generate a
It is important that ANDA applicants assess the potential of
greater immune response as compared to the RLD, Lovenox.126 In the case of enoxaparin, an
immune response can be generated that may lead to a known adverse event, thrombocytopenia.
There is evidence that the immune response resulting in thrombocytopenia is stimulated by the
this response has been extensively investigated and
active ingredient. The pathogenesis of
saccharides to
involves a critical step of association of enoxaparn (or other L WMHs) oligo
platelet factor 4 (PF4) via a non-specific (sequence independent) electrostatic

interaction
127 Therefore, a demonstraprimarily based on oligosaccharide chain length and charge density.
the generic drug product's enoxaparin to Lovenox's

enoxaparin is a strong indication that the generic enoxaparin would not differ from Lovenox with
respect to its immunogenicity. Nonetheless, immune responses are very complex and even when
they are stimulated by the active ingredient (as is the case with enoxaparin sodium), they may be
influenced by impurities or other substances in the product that may modify the immune
tion of equivalence of
molecular diversity of
response to the active ingredient. Given the safety profile of Lovenox, we believe that, in
the generic drug product's enoKaparin to Lovenox's
addition to demonstrating sameness of
enoxaparn, sponsors should submit a comparative assessment of their generic enoxaparin and
125 Response to ISTA at 11.
126 ANA applicants are not only required to demonstrate active ingredient sameness, but also to assure that the
the drug are
methods used in, or the facilities and controls used for, the manufacture, processing, and packaging of
this
the drg (see section II.A of
adequate to assure and preserve the identity, strength, quality and purity of
response).
the Structural Requirements for a carbohydrate based anticoagulant

with a reduced risk of inducing the immunological tye of heparin-associated thombocytopenia," Thrombosis and
Haemostasis 74:886-892; and Newman, P.M., Swanson, R.L" and Chong B.H. (1998), "Heparin-induced thrombocytopenia: IgG binding to PF4-heparin complexes in the fluid phase and cross-reactivity with low molecular weight
127 Greinacher, A. (1995), "Characterization of
hepari and heparinoid." Thrombosis and Haemostasis 80:292-297.
41
Docket No. FDA.2003-P-0273

Lovenox for potential impurities that may have an adverse impact with respect to immunogenicity. Based on review of available data, we have determined that it is possible, by satisfying the
five criteria and (as a conservative measure) by conducting in vitro and in vivo assays to address
impurities, to provide scientifically appropriate assurance that the risk of immunogenicity due to
potential impurities in the generic enoxaparin wil not be greater than that of Lovenox. In such
cases, clinical testing to demonstrate safety and effectiveness is not necessary for enoxaparin
sodium injection (see also sections VLC. and VLF.). We expect that a generic enoxaparin that
meets the requirements for ANDA approval wil be therapeutically equivalent to and can be
substituted for Lovenox with the expectation that it wil produce the same clinical effect and
safety profile as Lovenox.
F. Although FDA Regularly Takes
Note of the Actions of Other National or Interna-
tional Regulatory Authorities, Those Actions Do Not Constrain Our DecisionMaking

You state that the European Committee for Medicinal Products for Human Use held its plenary
meeting in June 2006 (A ventis August 25, 2006, Comment at 2). You also state that the meeting
report reflects that the Co-coordination Group for Mutual Recogntion and Decentralised
Procedures has determined that LMWHs should be considered "biological medicinal products."
(Aventis August 25,2006, Comments at 2). You state that this means that applicants for generic
LMWHs in the European Community may not seek approval of their products as generic
medicinal products (A ventis August 25, 2006, Comments at 2). Instead, a company seeking
approval of a generic LMWH wil need to seek approval as a "similar biological medicinal
product," which you state requires the submission of far more data than is submitted in an
ANDA (A ventis August 25, 2006, Comments at 2). You also state that the European Medicines
Agency's (EMEA's) Committee for Medicinal Products for Human Use published a draft
"Concept Paper on Similar Biological Medicinal Products Containing Low Molecular Weight
Heparins - (Non) Clinical Issues" (the "Concept Paper") (Aventis March 2,2007, Comments at
2).128 You also submitted the "EMEA, Committee for Medicinal Products for Human Use
(CHMP): Guideline on Non-Clinical and Clinical Development of Similar Biological Medicinal
Products Containing (LMWHsJ" (EMEA Guideline) (Aventis April 14, 2009, eomments at 1).
The EMEA Guideline recommends in vitro and in vivo pharacodynamic studies, in vivo
toxicological studies, and clinical studies (Aventis April 14,2009, Comments, Attachment). The
EMEA Guideline also recommends methods for the detection and monitoring of HIT in the
clinical triaL.
128 You state that according to the draft Concept Paper, the EMEA indicates that the assessment of applications for
the LMWHs
diffcult for several reasons including (1) the limited physicochemical characterization of
LMWHs is
the molecules and the limited knowledge about the qualitative and quantitative
due to the high complexity of
contrbution to safety and effcacy of each fraction and (2) the fact that while the kinetics ofLWMH are based upon
pharmacodynamic measurements, there is no demonstrated quantitative correlation between pharmacodynamics and
clinical efficacy (Aventis March 2,2007, Comments at 2). You state that for these reasons, the EMEA indicates that
classical bioequivalence studies are not suffcient to establish therapeutic equivalence between LMWHs (Aventis
these diffculties, the draft Concept Paper recommends
March 2,2007, Comments at 2). You state that, in light of
drafting a guideline on the (non) clinical aspects of the development and assessment of similar biological products
containng LMWHs (Aventis March 2, 2007, Comments at 2). You ask that we give the European decision
consideration as we evaluate your petition (Aventis March 2,2007, Comments at 3). The EMEA requested public
comment on the draft Concept Paper. We have considered the draft Concept Paper.
42
Although FDA regularly takes note of the actions of other national or international regulatory
authorities, those actions do not constrain our decision-making. The European regulatory
authority has different standards and procedures for the review and approval of drugs and
biological products. Moreover, although we generally concur with the EMEA conclusion of
possible limitations regarding
(1) physicochemical characterization ofLMWHs, (2) biological
and biochemical assays, and (3) demonstration of comparable kinetics based upon thephara-
codynamic measurements, we would not draw a conclusion of active ingredient sameness for
physicochemical properties (section lILA),
enoxaparin based solely upon equivalence of
in vivo
of
biological and biochemical assays (section IILD), and equivalence
equivalence of
pharacodynamic profiles (section III.E).
Rather, we can draw a conclusion of active ingredient sameness based upon five criteria, two of
which were not specifically considered by the EMEA Guideline, including equivalence of
heparin source material and mode of depolymerization (section IILB) and equivalence of
disaccharide building blocks, fragment mapping, and sequences of oligosaccharide species
(section IILe). As explained earlier in this response, we have concluded, based on the current
scientific evidence, that if an ANDA applicant meets the five specified criteria, this information
enables us to conclude that a generic drug product's enoxaparin is the same active ingredient as
Lovenox's enoxaparin. In such instances, an ANDA applicant would meet the requirement for
active ingredient sameness under the Act and FDA regulations.
Furter, we note that the EMEA Guideline has set forth guidelines for L WMH products that
contain a similar (as opposed to the same) active ingredient as that contained in another already
marketed LMWH product. Because the proposed product in Europe wil contain an active
ingredient that is similar (as opposed to the same), it may exhibit important differences from the
active ingredient in the already marketed LMWH product. Accordingly, there might be uncertainties regarding the safety and effectiveness of the proposed similar product. Thus, sponsors
under the EMEA framework are expected to provide clinical studies showing comparable
effectiveness to the proposed similar LMWH product as well as clinical data showing comparable safety, including with respect to HIT.
This contrasts with the approach taken by FDA. In this petition response, the Agency has set
forth the basis upon which we can conclude that a generic enoxaparin contains the same active
ingredient as the RLD, Lovenox. Because an ANDA for generic enoxaparin must contain
sufficient data demonstrating that the active ingredient is the same (as opposed to similar in the
EMEA framework) as Lovenox's enoxaparn, and because the Agency wil carefully evaluate
impurities in the generic product, particularly with respect to their adjuvant effect on immunogenicity, there is no underlying scientific need for the ANDA sponsor to duplicate clinical
studies to demonstrate safety (such as with respect to HIT) and effectiveness.
Furthermore, although the EMEA Guideline describes the need for clinical studies in order to
demonstrate comparable effectiveness to the proposed similar LMWH, the Agency notes the five
criteria described above in section III of this response are more sensitive to differences between
two enoxaparin products than the clinical studies recommended in the EMEA Guideline. For
example, a double blind clinical study showed that the rating of deep-vein thrombosis after
43
orthopedic surgery was equivalent for two LMWHs: enoxaparin and tinzaparin.129 However, as
discussed under section III.B of this response, despite such evidence of equivalence of clinical
effectiveness, enoxaparin and tinzaparin would not be considered to have the same active
ingredient, given that they are manufactured via different modes of depolymerization, 130 have
different anti-lla/anti-Xa ratios, 13 have different proportions of sulfated and non-sulfated !i4,5 uronate strctures at the non-reducing end of
the oligosaccharide chains,132 and have different
pharmacokinetics,133 among other differing characteristics.
We considered whether we should expect ANDA applicants of generic enoxaparn.to conduct

any studies recommended in the EMEA's guideline other than the ones we are recommending in
section III of this response. Based on our scientific experience and expertise, and relevant
scientific information, we have determined that no other studies are needed for approval of a
generic enoxaparin.
VII. CONCLUSION
For the reasons stated above, we conclude that the following five criteria are sufficient to
demonstrate sameness of the enoxaparin active ingredient:
. Equivalence of physicochemical properties
. Equivalence of
heparin source material and mode of depolymerization
.. Equivalence in disac~haride building blocks, sequence of oligosaccharide species, and
fragment mapping
. Equivalence in biochemical and biological assays
. Equivalence of in vivo pharmacodynamic profile
Meeting these specified active ingredient sameness criteria wil ensure that the molecular
the generic drug product's enoxaparin and Lovenox's enoxaparin wil be equivalent,
diversity of
including with respect to the 1,6 anhydro ring structure. Equivalent molecular diversity demonstrates enoxaparin sameness. The Act and FDA regulations require ANDA applicants to
demonstrate (among other things) that the generic drug product's active ingredient is the same as
the RLD's active ingredient and to demonstrate that the methods used in, or the facilities and
controls used for, the manufacture, processing, and packing of the drug are adequate to assure
Deep Vein Thrombosis after Hip Replacement: Comparison between
Two Low-Molecular Heparins, Tinaparin and Enoxapann," Thrombosis andHaemostasis 81: 22-25.
129 Planes, A. et a!. (1999), "Prevention of
130 Whereas enoxaparin is manufactured by chemical alkaline -elimination, tinzaparin is manufactured via
enzymatic -elimination.
131 For tinzaparin anti-lla/anti-Xa ratios varies between 1.5 and 2.5 (see footnote 33), and for enoxaparin this ratio
varies between 3.3 and 5.3 (see footnote 54 ).
132 This is because in the mode of depolymerization by enzymatic -elimination, cleavage takes place exclusively in
heparin polysaccharide chains at sites where the disaccharide unit has the 2-0-sulfoiduronic acid structure, whereas
in the mode of depolymerization by chemical (alkaline) -elimination, cleavage occurs without preference for the
presence or absence ofa 2-0-sulfo group in the iduronic acid structure. See Linhardt, RJ., Gunay, N.S. (1999),
"Production and Chemical Processing of Low Molecular Weight Heparin," Semin Thromb Hemost 25 S3: 10.
133 Eriksson, B.I. et.a!., (1995), "A comparative study of three low-molecular weight heparins (LMWH) and
unfractionated heparin (UH) in healthy volunteers," Thrombosis and Haemostasis 73: 398-401.
44
that an ANDA applicant use a

manufacturing process equivalent to Aventis's manufacturing process. Insofar as an equivalent
manufacturing process for enoxaparin could be interpreted only to be one in which every step of
the manufacturing process, including process conditions, are identical to those of Aventis's
manufacturing process for Lovenox, your request is denied. We also are denying your requests
that we require an ANDA applicant to completely characterize enoxaparin by isolating, purifying, and sequencing each of its unique polysaccharide chains and determining their relative
abundance or to conduct clinical trials to establish the safety and effectiveness of its product.
and preserve its identity, strength, quality, and purity. You ask
Sincerely,
4; c~h~
Douglas Throckmorton, M.D.

Deputy Director
Center for Drug Evaluation and Research
45

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('l

DEPARTMET OF HETH &. HUMA SERVICES

Re: Docket No. FDA-2003-P-0273

Dear Mr. Safir and Mr. Cunningham:

Lovenox (enoxaparin sodium injection) (hereafter, "generic enoxaparin")

the Federal Food, Drug, and Cosmetic Act (the Act).

Docket No. FDA-2003-P-0273

the RLD (i.e., Aventis's Lovenox). The crux of

ered. Lovenox, a naturally sourced drug product, has some degree of

Docket No. FDA-2003-P-0273

between 15 percent and 25 percent of its poly( oligo )saccharides.

Pharmaceutical Science (including the Office of

Biotechnology Products), and the

New Drug Quality Assessment, and the Office of

Docket No. FDA-2003-P-0273

immunogenicity. It is important that ANDA applicants assess the potential of

product to generate a greater immune response as compared to the RLD, Lovenox.

. Outpatient treatment of acute DVT without pulmonary embolism

certification benefits described in section 505 of

submitted by ANA applicants under section 505(j)(2)(A)(vii) of

those certifications, Aventis

Docket No. FDA-2003-P-0273

unstable angina and non-Qwave myo-

cardial infarction (MI)

aged medically or with subsequent percutaneous coronary intervention (PCI).I i

have made porcine tissue the principal source of

heparin. Heparin products currently used in the

induced thrombocytopenia (HIT), which is a serious antibody-mediated reaction resulting from

polysaccharide chains into smaller oligosaccharide

Docket No. FDA-2003-P-0273

Products with Therapeutic Equivalence Evaluations (Orange Book).

LMWHs, as noted above, are manufactured through depolymerization ofheparn.17 Both

respective RLDs. The recent severe anaphylactoid reactions reported following

this contamination, the u.s. Pharmacopeia (USP) has revised its

Biotechnology 26: 669-675. As a result of

Docket No. FDA-2003-P-0273

heparin is important for understanding the molecular

the molecular diversity of

diversity of LMWH. Heparin is a mixture of

5 01 oSOi OOSOi ~C00-2' OOSOi CO2 00SOi

~. :'. '.j" ." ".~ "0; .J

I ~SOj __~::o I ~H ~c I ~ '-AC I ~H ~o I

heparin is the polydisperse chain length of

the molecular diversity of

molecular weight of ~ 15,000 Da. The polydispersity of

between approximately 1.1 and 1.6.19

heparin's molecular diversity derives from the diversity of disaccharide

The second dimension of

chains. The diversity of disaccharide units (constituent building blocks of polysaccharide

two different uronic acid components (iduronic or

26 A disaccharide is a carbohydrate composed of two monosaccharides.

Docket No. FDA-2003-P-0273

sulfated.30 The molecular diversity of

having a 4,5 delta (4,5)-uronate structure at the non-reducing end of

cleavage using nitrous acid results in oligosaccharide fragments with a 2,5-anydro-D-mannitol

the benzyl ester derivative of

heparin," whereas for daltepari it

Docket No. FDA-2003-P-0273

the drug product. Instead, an ANDA applicant relies on the Agency's