LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-1
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-2
`IPR2016-00379
`
`

`
`First published in 1988 by
`ELLIS HORWOOD LIMITED
`Market Cross House , Cooper Street,
`Chichester, West Sussex, P019 lEB, England
`The publisher's colophon is reproduced from James Gillison's drawing of the ancient Market Cross,
`Chichester.
`
`Distributors:
`Australia and New Zealand:
`JACARANDA WILEY LIMITED
`GPO Box 859, Brisbane , Queensland 4001, Australia
`Canada:
`JOHN WILEY & SONS CANADA LIMITED
`22 Worcester Road, Rexdale, Ontario, Canada
`Europe and Africa:
`JOHN WILEY & SONS LIMITED
`Baffins Lane, Chichester, West Sussex, England
`North and South America and the rest of the world:
`Halsted Press: a division of
`JOHN WILEY & SONS
`605 Third Avenue, New York, NY 10158, USA
`South-East Asia
`JOHN WILEY & SONS (SEA) PTE LIMITED
`37 Jalan Pemimpin # 05-04
`Block B , Union Industrial Building, Singapore 2057
`Indian Subcontinent
`WILEY EASTERN LIMITED
`4835/24 Ansari Road
`Daryaganj, New Delhi 110002, India
`© 1988 S. G. Allenmark/Ellis Horwood Limited
`
`British Library Cataloguing In Publication Data
`Allenmark, S. G . (Stig G.) , 1936-
`Chromatographic enamtioseparation.
`l . Chromatography
`I. Title
`543'.089
`Library of Congress Card No. 88-1092
`ISBN 0-84312-988-6 (Ellis Horwood Limited)
`ISBN 0-470-21080-X (Halsted Press)
`Typeset in Times by Ellis Horwood Limited
`Printed in Great Britain by Hartnolls, Bodmin
`
`COPYRIGHT NOTICE
`All Rights Reserved. No part of this publication may be reproduced , stored in a retrieval system, or
`transmitted , in any form or by any means, electronic, mechanical, photocopying, recording or otherwise,
`without the permission of Elhs Horwood Limited, Market Cross House, Cooper Street, Chichester, West
`Sussex, England.
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-3
`IPR2016-00379
`
`

`
`Table of contents
`
`Preface . ... . .......... . ........ . ............... . ....... 9
`
`List of Symbols and Abbreviations ................ . ... . ......... 11
`
`1 Introduction ..... .. .... .. .. .. . ....................... 13
`Bibliography .................... . ............. . ..... . 18
`References ..... . ....................... ... .... . . .. .. 18
`
`2 The development of modern stereochemical concepts
`2.1 Chirality and molecular structure ......................... 19
`2.1.1 Molecules with asymmetric atoms ...................... 19
`2.1.2 Other types of chiral molecular structures ..... . ........... 20
`2.2 Definitions and nomenclature .......... . ................ 23
`Bibliography ........................... . ...... . .. . . .. 26
`References . ....... . ................ .... .......... .. . 26
`
`3 Techniques used for studies of optically active compounds
`3.1 Determination of optical or enantiomeric purity ............... 27
`3.1. l Methods not involving separation ... .. ................. 27
`3.1. l.1 Polarimetry ................................. 27
`3.1.1.2 Nuclear magnetic resonance .. .. . .. .. . ...... . ..... . 29
`3.1.l.3 Isotope dilutio n .................. . ..... .. ..... 31
`3.1.l.4 Calorimetry .... . ... .. . .. ... . ................ 33
`3.1.1.5 Enzyme techniques ................... . ......... 33
`3.1. 2 Methods based on separation ......................... 34
`3.2 Determination of absolute configuration .................... 35
`3.2.l X-Ray crystallography with anomalous scattering ...... . .. ... 36
`3.2.2 Spectroscopic (ORD , CD) and chromatographic methods
`based on comparison ......... .. ..... .. ............ 37
`Bibliography ................... .. .. ...... .... ...... . .40
`References .. . . ... ............... . ................... 40
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-4
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-5
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-6
`IPR2016-00379
`
`

`
`8
`
`Table of contents
`
`Bibliography .................. . • ........ .. ..... . .. . .. 188
`References .. .... ... .. . .. .... . . ... ... .. .. .. . . .. .. .. . 188
`
`9 Preparative scale enantioseparations- need, progress and problems . . .. 192
`Bibliography . . .... . ..... . ............ . ... .. . .... .... 199
`References ... .. . . . . ... .. . ... . .... .. .... . .. . . .. ..... 199
`
`10 Future trends
`10.l New detector systems . .. .. .. .... . .. .. .... . .... . ... . .. 200
`10.2 Column improvements . . . . ... . .... . ... .. .... . ... . .... 203
`10.3 Supercritical fluid chromatography . . . . .... . . .. .. . ........ 206
`Bibliography .. . .... ... . .. .. ... ... .. ....... .. . . . ..... 206
`References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206
`
`11 ~xperimental procedures for the synthesis of chiral sorbents
`11.1 Techniques for the preparation of chiral sorbents by
`derivatization of polysaccharides . .. .... ... .. .. . .. .. . . .. . 208
`11.1.1 Preparation of microcrystalline cellulose triacetate (MCTA) .... 208
`11.1 .2 Preparation of silica coated with cellulose triacetate .. .. . . .. . 209
`11.1.3 Preparation of silica coated with cellulose triphenylcarbamate ... 209
`11.2 Polymerization procedures used to obtain chiral synthetic
`polymer materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 209
`11.2.1 Preparation of poly[(S)-N-acryloylphenylalanineethyl ester] ... . 209
`11.2.2 Preparation of polycellulose(triphenylmethyl methacrylate) ... . 211
`11.3 Techniques used for the binding of chiral selectors to silica .. .. . ... 211
`11 .3.1 Preparation of 3-glycidoxypropyl-silica ... . ... ........ . . . 212
`11 .3.2 Large-scale preparation of (R)-N-(3,5-dinitrobenzoyl)-
`phenylglycine covalently silica-bound sorbent . .. .. .. . .. . .. 212
`11 .3.3 Preparation of (S)-(- )-~-N-(2-naphthyl)leucine .. .. .. ...... 213
`11 .3.4 Hydrosilylation of (R)-N-(10-undecenoyl)-~-(6,7-dimethyl-
`1-naphthyl) isobutylamine . ... ... . .. . . . . . . .. ... ... . . 213
`11.3.5 Preparation of silica-bonded (S) -1-(~-napththyl)ethylamine .... 213
`11.3.6 Preparation of silica-bound polyacrylamide and
`polymethacrylamide . . . .. . .... . ...... . ......... . . . 214
`Bibliography .... .. . ... ... .. .. . .. . . .. .. .. . .. .. . ... .. . 214
`References .... . ... . ... . ... ... ... . .. . ... . . .. ... . . ... 214
`
`Appendix: Commercial suppliers of chiral columns for GC and LC . . ....... 216
`
`Index . .. .. ....... .. .... . .... .. ...... .. . . ...... .. .. . . . 218
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-7
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-8
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-9
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-10
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-11
`IPR2016-00379
`
`

`
`46
`
`Modern chromatographic separation methods
`
`[Ch. 4
`
`It should be noted that for ex-values close to 1, small changes in ex will result in large
`changes in R,. Accordingly , a change in ex from 1.1to1.2 means roughly a doubling of
`Rs, owing to the influence on the (ex - 1 )/ex factor. Equation ( 4. 7) is often simplified
`by the introduction of the concept Nem the effective number of theoretical plates.
`Because of the definition Neer= 16[(tR -t0 )1w]2 , cf. Eq. (4.1) , it can be shown that
`Eq . ( 4. 7) can be rearranged into the simpler form
`
`-
`ex-1
`Rs =~ \!Neff
`
`(4.8)
`
`where the factor k2f(l + k;) is incorporated into Nerr: Neer= N[k'l (l + k ' )]2. From
`graphical evaluation of Eq. (4.8) at given R, values , the effective plate number
`required to obtain a certain resolution as a function of~ can be found. An illustrative
`example is given in Fig. 4.5. Thus, to obtain a 6cr resolution (R, = 1.5) an ex-value of
`1.05 will require 15700 effective theoretical plates, whereas the corresponding
`number needed for ex= 1.15 will be only 2110.
`
`INSTRUMENTATION
`4.2
`The basic instrumental requirements for GC as well as LC are rather simple: a system
`for mobile phase delivery to the column, an injection device for application of the
`sample, a separation column and a system for detection of the separated compo(cid:173)
`nents. In GC it is of course also essential to use a thermostatically controlled oven to
`contain the column , and separate temperature controls for the injection and
`detection units. The extraordinarily rapid development of GC and LC instruments ,
`however, has almost made instrument technology a science of its own. It is not the
`purpose of this chapter to give any detailed account of modern instrumentation, but
`merely to concentrate upon what is essential in this context with respect to separation
`of enantiomers. For further details on instrumentation the reader is referred to the
`exhaustive treatment by Poole and Schuette (see Bibliography).
`
`4.2.1 Gas chromatographic instrumentation
`The principal components in a gas chromatographic system are outlined in Fig. 4.6.
`The mobile phase or carrier gas should be inert under the conditions used and its
`selection will be dependent mainly on the type of detector used. Nitrogen, hydrogen,
`helium or argon are preferred. Hydrogen has the lowest viscosity of all , which means
`that it is used with advantage in long capillary columns where relatively high flow(cid:173)
`rates are required . This is sometimes of practical importance , e.g. when the
`temperature stability of the stationary phase is limited , since at a low column
`temperature retention times would otherwise be too long. The van Deemter curve,
`which relates the column plate height to the linear flow-rate of the mobile phase and
`from which the column efficiency can be optimized , is very different for hydrogen
`and nitrogen. As shown in Fig. 4.7 there is a much flatter minimum for hydrogen ,
`which leads to considerably higher column efficiency at high flow-rates.
`The carrier gas enters the column through the injector block and leaves through
`the detector system. There is a wide variety of injection modes, particularly for
`capillary columns. Normally , however, the injector block is kept at a temperature ca.
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-12
`IPR2016-00379
`
`

`
`Sec. 4.2)
`
`Instrumentation
`
`47
`
`log Neff
`
`5.0
`
`4.0
`
`3 .0
`
`1.00
`
`1.10
`
`1.20
`
`<l'
`
`Fig. 4.5- Plots of plate numbers vs. <:Y-values at different constant resolution values. Note the
`drastic increase in plate numbers needed to maintain resolution at very low <:Y-values.
`
`HiJ I F~\ I
`
`oc ~~f
`
`...____Detoot
`
`injector
`port
`
`column
`
`oven
`
`recorder / integrator
`
`Fig. 4.6- Essential features of a gas chromatograph.
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-13
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-14
`IPR2016-00379
`
`

`
`Sec. 4.2]
`
`Instrumentation
`
`49
`
`pack ed column
`
`open t ubular capillary colum ns
`
`<glas s o r steell
`
`0 <gla ss or qu ar t z)
`
`10 >2mm
`
`10< 0.5mm
`
`SCOT WCOT
`<supp ort - <wall (cid:173)
`coatedl coated )
`
`Fig. 4.8- Different column modifications used in gas chromatography.
`
`was introduced in 1979 for capillaries. The nature of the inner wall surface of the
`tubing is critical for maintenance of the stationary phase. The inner wall surface can
`be coated with the stationary phase liquid by various techniques to give a desired film
`thickness which then determines the capacity and retention ability of the column.
`Normally, the film thickness is 0.1-0.3 µm. When fused silica in particular is used , it
`is essential to 'immobilize' the stationary phase in some way, otherwise column
`bleeding will become a problem at higher temperatures. The various techniques used
`for this purpose are aimed at creating cross-links in the stationary phase polymer.
`Although the mechanisms of these reactions are not known in detail, it is likely that
`covalent bonding to the wall surface will take place to some extent. The main result is
`a column with very low bleeding at elevated temperatures, and high solvent
`resistance. Further, immobilization techniques make it possible to increase the film
`layer thickness considerably.
`Since resolution is of primary importance for enantiomer separation by GC,
`WCOT-columns are by far of greatest interest. Consequently, details of the wall
`coating and temperature stability of the stationary phase are important and will be
`considered further in later chapters.
`A variety of detectors have been developed for the monitoring of separated
`components in a GC column effluent. The most widely used belong to the ionization
`detector category. The principle applied is the measurement of change in electric
`conductivity caused by changes in ion currents generated in the detector. The flame
`ionization detector (FIO) is the most universally used. It meets all requirements of a
`good GC detector, such as high sensitivity, very good stability, fast response (1
`msec) , low dead volume (1 µl) and a broad linear response range. It operates by
`means of combustion of the effluent components in a hydrogen flame, thereby
`generating positive ions which cause an increase in the conductivity of the applied
`electric circuit.
`An even more sensitive and more selective ionization detector is the electron
`capture detector (ECO). As expected from the name, it operates through capture of
`electrons by the analytes, which means that certain demands are placed on the
`organic structure of these compounds. In ECO , the carrier gas molecules are ionized
`by a P-emitting radiation source. This ionization produces thermal electrons which
`generate a stable background current when subjected to a potential difference in the
`ECO-cell. When an electron-capturing compound is eluted from the column , it will
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-15
`IPR2016-00379
`
`

`
`50
`
`Modern chromatographic separation methods
`
`[Ch. 4
`
`diminish the background current and produce a signal on the recorder. The ECO can
`be said to operate in reverse mode to the FIO.
`The ECO, which was originally applied to high-sensitivity detection of haloge(cid:173)
`nated hydrocarbons, has proved excellent for use in combination with derivatization
`of amines, amino-acids, hydroxy-acids and similar compounds. Halogenated,
`notably perftuorinated, acylating reagents are used to introduce EC-sensitive groups
`into amino- and hydroxy-compounds by the formation of volatile amides and esters.
`The sensitivity of the EC-detector is highly dependent on the analyte strucure. A
`basic requirement is the ability of the compound to accommodate the negative
`charge produced by the electron capture. Accordingly, halogenated compounds,
`nitroaromatics, polycyclic aromatic hydrocarbons and conjugated carbonyl com(cid:173)
`pounds are among the structures giving a high detector response.
`
`4.2.2 Liquid chromatographic instrumentation
`Since a liquid mobile phase is used, the solvent delivery system forms an important
`part of a LC instrument. As high-efficiency columns usually produce a significant
`back-pressure, a high-pressure pump must be used to force the mobile phase through
`the column at a controlled flow-rate. The common instrumental set-up is shown in
`Fig. 4. 9. The sample is introduced as a solution by a syringe through an injection
`
`Mobile
`phase
`
`Delivery
`system
`
`Injection
`
`Column
`
`Recorder
`
`Integrator
`Data processing
`
`Fig. 4. 9 - Main components of a liquid chromatograph.
`
`usually a loop system. Normally, packed columns of 4-5 mm internal
`device -
`diameter and 15-25 cm length are used for analytical work. Many different types of
`column packing materials have been used, but the majority today are based on a
`silica support. If the LC modes based on molecular sieving effects are excluded, LC
`separations are completely dominated by the use of bonded phase materials, i.e.
`those in which the stationary phase is integrated with the (silica) support by covalent
`bonding. The chemistry of ligand coupling to silica particle surfaces has developed
`rapidly and a multiplicity of bonded phases for use in ion-exchange , reversed-phase
`and affinity chromatography is now available. The immobilization chemistry will be
`dealt with in further detail in connection with the presentation of chiral stationary
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-16
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-17
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-18
`IPR2016-00379
`
`

`
`Sec. 4.3) Separation of enantiomers by means of covalent diastereomers
`
`53
`
`derivatives (I and II) , obtained by reaction with ( + )-B, any contamination with
`(- )-B will be deleterious. The products (III and IV) produced with (- )-B form
`enantiometric pairs with the two main products (IV with I and III with II) and are
`therefore added to the corresponding peaks. The possible effect of this is best shown
`by an example. Let us assume that A consists of the (+)-form in 99% optical purity.
`The reagent ( + )-B is assumed to be 97% optically pure. By definition (see Section
`
`(±)-A
`
`(+)-8
`
`(+)-A-(+ )-8
`~ (-)-A-(+)-8
`main reaction
`
`~
`
`(+l-A-(-)-8
`
`(-)-A-(-)-8
`
`( I )
`
`(llJ J
`
`pairs of
`enant10-
`me r s
`
`( III )
`
`(IV)
`
`Scheme 4.2- Illustration of the effect of the use of a chiral derivatization reagent with less than
`100% optical purity. Because enantiomers cannot be separated on a non-chiral stationary
`phase, products I and IV give only one peak, as do II and Ill.
`
`3.1.1), there will be 99.5% of (+)-A, 0.5% of (-)-A, 98.5% of (+)-Band 1.5% of
`(- )-B present in the reaction mixture. Assuming complete reaction , the proportions
`of I-IV will then be: 98.0075% , 0.4925%, 1.4925% and 0.0075% respectively. Since
`I and IV are superimposed in the chromatogram, as well as II and III , the two peaks
`will show the proportions 98.015% and 1.985% , respectively. Therefore , if the
`optical purity of the derivatization reagent is not taken into consideration, A will be
`found to contain 98% of (+)-form, which corresponds to an optical purity of 96% , a
`considerable error relative to the true value.
`Secondly, the quantitative analysis relies upon the assumption that the reactions
`are complete . If this is not the case, differences in product yields may result in large
`errors. It is also very important to be sure that the reactions are not associated with
`racemization or epimerization. Compounds containing asymmetric carbon centres
`which could change configuration through carbanion or carbonium ion intermediates
`have to be treated particularly carefully as they may not possess the necessary
`configurational stability under conditions that are too basic or too acidic.
`Another factor of primary importance in chiral derivatization for the separation
`of diastereomers is the distance between the two chiral centres in the derivatives. As
`a general rule the centres should be as close as possible to each other in order to
`maximize the difference in chromatographic properties. Generally, three bonds
`separate the two centres and derivatives having distances exceeding four bonds are
`not often useful.
`For a detailed and comprehensive treatment of the chromatographic separation
`of diastereomers, the reader is referred to the recent book by Souter (see
`Bibliography).
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-19
`IPR2016-00379
`
`

`
`...
`
`54
`
`Modern chromatographic separation methods
`
`[Ch.4
`
`4.3.1 Gas chromatography
`It was shown as early as 1960 that it was possible to separate the L-alanyl derivatives
`of D,L-phenylalanine by GC as their N-triftuoroacetylated methyl esters (3). This led
`to the use of amino-acid derivatives as chiral N-derivatizing agents for the GC
`resolution of amino-acids as diastereometric dipeptide derivatives. A commonly
`used reagent is N-triftuoroacetyl-L-prolyl chloride, which may be prepared in high
`optical purity from L-proline, thanks to the resistance of this cyclic amino-acid to
`racemization. Other chiral reagents for the derivatization of the amino group in
`amino-acids are various 2-chloroacyl chlorides. These compounds can also be
`prepared from their corresponding amino-acids.
`For chiral derivatization of the carboxyl group, various optically active secondary
`alcohols have been used, such as 2-butanol or higher homologues. ( - )-Menthol has
`also been used for the preparation of diastereometric esters prior to GC (4).
`Hydroxy functions, as in alcohols or hydroxy-acids, are readily derivatized with
`(-)-menthyl chloroformate [5) to give diastereomeric esters or with ( + )- or (-)(cid:173)
`phenylethyl isothiocyanate [6) to yield carbamates.
`A number of other possibilities for chiral derivatization exist which have been
`found useful. A summary is given in Table 4.2, together with references to relevant
`original articles.
`
`4.3.2 Liquid chromatography
`Much of the early work in this area was, as in the case of GC, centred on amino-acids,
`particularly stereochemical analysis in peptide synthesis. A very comprehensive
`investigation of the separation of diastereomeric dipeptides on cation-exchangers
`was performed by Manning and Moore (56, 57). N-Carboxy-anhydrides of various
`amino-acids were used as chiral derivatization agents. The technique was later
`modified by the use of BOC-L-amino acid N-hydroxysuccinimide esters, followed by
`removal of the BOC group with triftuoroacetic acid [58). It was found that on
`reversed-phase LC of such dipeptides, the D,D- or L,L- forms were eluted before their
`D,L- or L,D- diastereomers. NMR investigations showed that in their most stable
`conformation only the latter have the hydrophobic side-chains cis-oriented. It was
`assumed that such an orientation would yield a more effective interaction with the
`hydrophobic stationary phase [59,60). Other very useful chiral derivatization re(cid:173)
`agents for amino-acids and RP-LC include the isothiocyanates of tetra-acetylglucose
`(GITC) and of triacetylarabinose (AITC) [61, 62). The latter are also suitable for
`amines [63] and amino-alcohols [64]. Optically active 1-phenylethyl isocyanate [65)
`has also been found useful for RP-LC of chiral amines. It has also been used, as has
`the 1-naphthyl analogue , for derivatization of alcohols to diastereomeric carbamates
`which may be separated by normal-phase LC [66, 67).
`A variety of diastereomeric amide and ester derivatives of chiral carboxylic acids
`have been used and methyl esters of L-amino-acids, such as L-phenylalanine [68] or
`L-phenylglycine [69], are readily available chiral reagents.
`A summary of the most useful chiral derivatization techniques in liquid chroma(cid:173)
`tography is given in Table 4.3.
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-20
`IPR2016-00379
`
`

`
`~
`~
`0
`(tl
`{/}
`
`References
`
`analyte
`Type of
`
`structure
`Product
`
`reagent
`Chiral derivatization
`
`Table 4.2 -Chiral derivatization reactions used in gas chromatography
`
`(16)
`
`Amines
`
`[17, 18]
`
`Amines
`
`[14-161
`[10, 12, 13)
`
`Amines
`Amino-.acids
`
`[ll]
`[ 11]
`
`[10]
`(7-9]
`
`Amines
`Amino-acids
`
`Amines
`Amino-acids
`
`Amines
`Amino-acids
`
`-N-~y~)~CF
`
`3
`
`R1
`
`H
`
`0
`
`6 5
`-N-C'D/......._c F
`
`H
`
`II
`0
`
`H
`3
`-N-C'cJ/ '-CF
`
`II
`0
`
`-~-Cv="
`
`/
`
`II
`0
`
`0
`
`II
`0
`
`2
`-N-C-CHCHMe
`
`-~-~ B-
`
`Cl
`I
`
`H
`
`II
`0
`
`CH2CHMe2, C6Hs)
`(R1 =CH3, CHMe2,
`and homologues
`chloride
`N-TFA-alanyl
`
`benzoyl)prolyl chloride
`N-(PentaHuoro-
`
`chloride
`N-TFA-Prolyl
`
`chloride
`Chrysanthemoyl
`
`Drimanoyl chloride
`
`chloride
`2-Chloroisovaleryl
`
`-N-C-R
`II *
`0
`
`-NH2-
`
`transformation
`Functional group
`
`3
`'<
`r::r
`"'
`'"I
`~
`3
`= -
`cs·
`~ = =
`= 0 -
`cs·
`=
`= '"I
`
`-
`
`'C
`~
`r:J'J
`
`-n
`
`< = ;:;" = -
`~ = = "'
`
`"'
`3
`0
`~
`..,
`,,.
`;·
`Q.
`
`-~
`
`
`
`Vl
`Vl
`
`'"I
`~
`
`
`
`0
`
`0
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-21
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-22
`IPR2016-00379
`
`

`
`-.J
`VI
`
`"'
`.,
`0 = ~
`.,
`ir
`c.
`ii' =
`
`
`-~
`-
`
`~
`
`lj)
`~
`0
`t'l
`0
`
`~
`~
`
`~
`
`....
`
`0
`
`=
`.,
`
`"O
`~
`r;n
`
`~
`~
`ri
`(b
`(I')
`
`....
`"'
`=
`= ~ =
`"'
`.,
`=
`~ = = = -
`s·
`= -s· =
`
`[41)
`[29, 38-40)
`(27' 28, 34--37)
`
`Keto-acids
`a-Hydroxy-acids
`Amino-acids
`
`-c,o...-CH-CH3
`H
`0
`
`~1
`
`(R1=n-CrC61 Me2CH, Me3C)
`2-Alkanols
`
`(6, 33]
`
`Hydroxy-acids
`
`(22, 30--32)
`(22]
`
`Alcohols
`Hydroxy-acids
`
`[1~21]
`
`(OH)
`Amphetamines
`
`[11]
`[11]
`
`(11]
`
`References
`
`Alcohols
`Hydroxy-acids
`
`Hydroxy-acids
`
`analyte
`Type of
`
`6 5
`-o-c,.....N' CH-C H
`
`CH3
`I
`
`0
`II
`
`H
`
`09
`
`-o-c-...0
`
`II
`
`OCH3
`
`-o-c-?-C6HS
`
`~ yF3
`
`-o-cy=,
`
`/
`
`II
`0
`
`OB
`
`-0-C
`II
`
`structure
`Product
`
`Table 4.2 (contd.)
`
`-C02R
`*
`
`-C02H -
`
`isocyanate
`1-Phenylethyl
`
`-OH--0-C-N-R
`*
`
`II
`0
`
`H
`
`(-)-Menthyl chloroformate
`
`-OH--0-C-OR
`*
`
`II
`0
`
`MTPA-Cl
`
`chloride
`Chrysanthemoyl
`
`chloride
`Drimanoyl
`
`reagent
`Chiral derivatization
`
`transformation
`Functional group
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-23
`IPR2016-00379
`
`

`
`.;..
`
`n ::r
`
`Cll
`Q.
`ft) -
`Q
`::r
`a
`=
`= -
`s·
`"O =
`..,
`;:;·
`::r
`=
`"O
`(1CI ..,
`= -Q
`a
`..,
`::r
`=
`..,
`::: Q
`
`Q
`
`!')
`
`ft)
`Q.
`
`ft)
`Cll
`
`[3, 53]
`
`Amino-acids
`
`[52)
`
`Amino-acids
`
`{46-51]
`[45)
`[4, 42-44)
`
`References
`
`Other acids
`a-Hydroxy-acids
`Amino-acids
`
`analyte
`Type of
`
`-C-N-CH-CO Me
`II
`0
`
`2
`
`H
`
`I
`R
`
`2
`-C-N-CH-CH -CHMe
`
`2
`
`H
`
`yH3
`
`II
`0
`
`0 -2
`
`II -c,o
`
`structure
`Product
`
`00
`VI
`
`Table 4.2 ( conrd.)
`
`*
`'
`/C=O-/C=N-0-R
`
`'
`
`1-phenylethylhydrazine
`( + )-2,2,2· Trifluoro-
`
`/
`CsO-C=N-N-R
`*
`......
`
`/
`'
`
`H
`
`methyl esters
`Amino-acid
`
`pentylamine
`( + )-4-Methyl-2-
`
`( -)-Menthol
`
`-C-N-R
`*
`II
`0
`
`H
`
`-CO H -
`
`2
`
`reagent
`Chiral derivatization
`
`transformation
`Functional group
`
`r
`
`[55)
`
`[54)
`
`Ketones
`
`6 s
`C=N-N-CH-C H
`
`H
`
`,,
`'
`
`CF3
`
`I
`
`~C=N,0~ Carbohydrates
`
`0-(-)-Menthylhydroxylamine
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-24
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-25
`IPR2016-00379
`
`

`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-26
`IPR2016-00379
`
`

`
`<
`0
`n
`....
`0
`= 1;11
`=
`ft>
`3
`'<
`~
`1;11
`
`ft> .,
`0 3
`ft> .,
`1;11 ...
`&r
`= ii' = -Q.
`
`1;11
`
`ft>
`
`ft> .,
`3
`... :s·
`= =
`ft> =
`....
`:s· =
`...
`=
`=
`.,
`
`0
`
`"Cl
`ft>
`r:ll
`
`[84)
`
`[83]
`
`(82]
`
`[81]
`
`acids
`Carboxylic
`
`acids
`Carboxylic
`
`acids
`Carboxylic
`
`acids
`Carboxylic
`
`0
`-¥-N-?H-C02CH3
`
`CH2C6H5
`
`H
`
`Methyl phenylalaninate
`
`0
`u
`-c-~-cH-@-N02
`
`CH3
`I
`
`1-( 4-Nitrophenyl)ethylamine
`
`-C-~-CH_g
`
`CH3
`I
`
`0
`11
`
`1-(1-Naphthyl)ethylamine
`
`0
`N
`-c-~-cH-@
`
`CH3
`I
`
`1-Phenylethylamine
`
`•
`
`H
`
`II
`0
`
`-CO H--C-N-R
`
`2
`
`~
`~
`!"l
`(1)
`(/.)
`
`References
`
`analyte
`Type of
`
`structure
`Product
`
`reagent
`Chiral derivatization
`
`lransformation
`Funclional group
`
`Table 4.3 (contd.)
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-27
`IPR2016-00379
`
`

`
`62
`
`Modern chromatographic separation methods
`
`[Ch.4
`
`BIBLIOGRAPHY
`E. ·Gil-Av and D. Nurok, Resolution of Optical Isomers by Gas Chromatography of Diastereoisomers,
`Adv. Chromatog., 1972, IO, 99.
`L. R. Snyder and J. J. Kirkland, Introduction to Modern Liquid Chromatography, Wiley, New York,
`1974.
`K. Blau and G. S. King, Handbook of Derivatives for Chromatography, Heyden, London, 1978.
`C. F. Poole and S. A. Schuette, Contemporary Practice of Chromatography, Elsevier, Amsterdam, 1984.
`R. W. Souter, Chromatographic Separation of Stereoisomers, CRC Press, Boca Raton , 1985.
`REFERENCES
`[I] L. S. Ettre and A . Zlatkis (eds.), 75 Years of Chromatography. A Historical Dialogue, Elsevier,
`Amsterdam, 1979.
`[2] M. Novotny, Anal. Chem., 1981 , 53, 1294A.
`[3] F. Weygand, B. Kolb , A. Prox, M. Tilak and I. Tomida, z. Physiol. Chem., 1960, 322, 38.
`(4] B. Halpern and J. W . Westley, Chem. Commun., 1965 , 421.
`[5] R. G. Annett and P. K. Stumpf, Anal. Biochem., 1972, 43, 515.
`(6] M. Hamberg, Chem. Phys. Lipids, 1971 , 6, 152.
`(7] J. W. Westley and B. Halpern, in Gas Chromatography, C. L.A. Harbourn (ed. ), Inst. of Petrol. ,
`London 1968, p. 119.
`(8] B. Halpern and J. W. Westley, Chem. Commun., 1965 , 246.
`(9] S. Lande and R. A. Landowne, Tetrahedron, 1966, 3085.
`(10) B. Halpern and J. W. Westley, Chem. Commun., 1966, 34.
`(11) C. J. W. Brooks, M. T. Gilbert and J. Gilbert, Anal. Chem . 1973, 45, 896.
`[12) B. Halpern and J. W. Westley, Biochem. Biophys. Res. Commun ., 1965, 19, 361.
`[13] B. Halpern and J. W. Westley, Tetrahedron Leu., 1966, 2283.
`[14) J . W. Westley and B. Halpern, Anal. Chem., 1968, 40, 2046.
`[15) R. W. Souter, J. Chromatog., 1975 , 108, 265.
`(16) B. L. Karger, R. L. Stern and W. Keane, Anal. Chem., 1967, 39, 228.
`(17) S. B. Matin, M. Rowland and N. Castagnoli, Jr., J. Pharm . Sci., 1973, 62, 821.
`(18) S. B. Matin, S. H . Wan and J. B. Knight, Biomed. Mass. Spectrom., 1973, 4, 118.
`(19) J . A. Dale, D. L. Dull and H. S. Mosher, J. Org. Chem., 1969, 34, 2543.
`(20] J . Gal and M. M. Ames, Anal. Biochem., 1977, 83, 266.
`(21) J. Gal, J. Pharm. Sci., 1977, 66, 169.
`(22) J. W. Westley and B. Halpern, J. Org. Chem., 1968, 33, 3978.
`[23) S. Hammarstrom and M. Hamberg, Anal. Biochem., 1973, 52, 169.
`(24] C . J. W. Brooks and J. D . Gilbert, Chem. Commun., 1973, 194.
`[25) J. D. Gilbert and C. J . W. Brooks, Anal. Lett., 1973, 6, 639.
`[26] J. P. Guette and A. Horeau , Tetrahedron Leu. , 1965, 3049.
`[27) R. Charles, G. Fisher and E. Gil-Av, lsr. J. Chem., 1963, I, 234.
`(28] E. Gil-Av, R. Charles and G. Fisher, J. Chromatog., 1965, 17, 408.
`(29) S. Julia and J.M. Sans,J. Chromatog. Sci., 1979, 17, 651.
`[30) S. Hammarstrom, FEBS Lett., 1969, 5, 192.
`(31) M. Hamberg, Anal. Biochem., 1971 , 43, 515.
`(32] R . G . Annett and P. K. Stumpf, Anal. Biochem., 1972, 47, 638.
`[33) W. Pereira, Anal. Lett., 1970, 3, 23.
`(34) M. E. Bailey and H.B. Hass, J. Am. Chem. Soc., 1941, 63, 1969.
`(35) G . E. Pollock and V. I. Oyama, J. Gas Chromatog., 1966, 4, 126.
`(36) G . E . Pollock and A.H. Kawauchi , Anal. Chem., 1968, 40, 1356.
`(37) F. Raulin and B. N. Khare, J. Chromatog., 1973, 75, 13.
`(38) E . Gil-Av and D. Nurok, Proc. Chem. Soc., 1962, 146.
`(39) B. L. Karger, R. L. Stearn, H. C. Rose and W. Keane, in Gas Chromatography, A. B. Littlewood
`(ed.), Inst. of Petrol., London, 1967, p. 240.
`(40) J.P. Kamerling, M. Duran, G. J. Gerwig, D. Ketting , L. Bruinvis, J. F. G. Vliegenhart and S. K.
`Wadman, J. Chromatog., 1981 , 222, 276.
`(41] H. Schweer,J.Chromatog., 1982, 243, 149.
`(42) S. V. Vitt , M. B. Saporovskaya, J.P. Gudkova and V. M. Belikov, Tetrahedron Lett., 1965, 2575.
`[43] M. Hasegawa and I. Matsubara, Anal. Biochem., 1975, 63, 308.
`(44] R. Klein,J. Chromatog., 1979, 170, 468.
`[45] J . P. Kamerling, G. J. Gerwig, J. F. G. Vliegenhart, M. Duran, D. Ketting and S. K. Wadman , J.
`Chromatog., 1977, 143, 117.
`[46] I. Maclean , G . Eglinton, K. Douraghi-Zadeh, R. G. Ackman and S. N. Hooper, Nature, 1968, 218,
`1019.
`
`LOWER DRUG PRICES FOR CONSUMERS, LLC
`Exhibit 1012-28
`IPR2016-00379
`
`

`
`Ch. 4]
`
`References
`
`63
`
`[47] R. E. Cox, J. R. Maxwell, G. Eglinton and C. T. Pillinger, Chem. Commun., 1970, 1639.
`[48] R. G. Ackman, R. E. Cox, G. Eglinto n, S. N. Hooper and J . R. Maxwell, ] . Chromatog. Sci., 1972,
`IO, 392.
`[49] A . Murano, Agr. Biol. Chem., 1972, 36, 2203.
`(50] M. Miyakado , N. Onno , Y. Okuno, M. Hirano, K. Fujimoto and H. Yoshioka, Agr. Biol. Chem.,
`1975, 39, 267.
`(51] M. Horiba, H . Kitaharo , H . Takahashi , S. Yamamota, A. Murano and N. Oi, Agr. Biol. Chem.,
`1979, 43, 2311.
`(52] B. Halpern, L. F. Chew and J. W. Westley, Anal. Chem., 1967, 39, 399.
`(53] F. Weygand, A. Prox, L. Schmidhammer and W. Konig , Angew. Chem., 1963, 7S, 282.
`(54] W. E . Pereira, M. Solomon and B. Halpern, Aust. J. Chem., 1971, 24, 1103.
`(55] H. Schweer, J. Chromatog., 1982, 243, 149.
`(56] J. M. Manning and S. Moore, J. Biol. Chem., 1968, 243, 5591.
`(57] J.M. Manning, J. Biol. Chem., 1971 , 246, 2926.
`(58] A . R. Mitchell, S. B. H. Kent , I. C. Chu and R . B. Merrifield, Anal. Chem., 1978, SO, 637.
`(59] E . P. Kroeff and D. J. Pietrzyk, Anal. Chem., 1978, SO, 1053.
`(60] E. Lundanes and T . Geibrokk, J. Chromatog., 1978, 149, 214.
`(61] N. Nimura, H. Ogura and T . Kinoshita, J. Chromatog., 1980, 202, 375.
`(62] T. Kinoshita , Y . Kasahara and N. Nimura, J. Chromarog. , 1981 , 210, 77.
`(63] A . J. Sedman and J . Gal, J. Chromarog. , 1983, 278, 199.
`[64] N. Nimura, Y. Kasahara and T. Kinoshita, J. Chromarog., 1981, 213, 327.
`(65] J. A. Thompson, J. L. Holtzman, M. Tsuru and J. L. Holtzman,]. Chromatog., 1982, 238, 470.
`[66] W. H . Pirkle and J. R . Hauske , J. Org. Chem., 1977, 42, 1839.
`(67] W. H. Pirkle and K. A. Simmons and C. W. Boeder, J. Org. Chem., 1979, 44, 4891.
`(