PCT
`WORLD INTELLECTUAL PROPERTY ORGANIZATION
`International Bureau
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`WO 00/55131
`(51) International Patent Classification 7 :
`C07D 207/12, A61K 31/40
`
`Al
`
`(11) International Publication Number:
`
`(43) International Publication Date: 21 September 2000 (21.09.00)
`
`(21) International Application Number:
`
`PCT/DK00/00104
`
`(22) International Filing Date:
`
`13 March 2000 (13.03.00)
`
`(30) Priority Data:
`PA 1999 00363
`
`15 March 1999 (15.03.99)
`
`DK
`
`(71) Applicant: NOVO NORDISK A/S [DK/DK]; Novo Alle,
`DK-2880 Bagsvaerd (DK).
`
`(72) Inventors: JESSEN, Claus, Ulrich; Fossglirdsvej 4, DK-2720
`Vanl0se (DK). WIEDE, Petra; GodthSbsvej 119, 4. th.,
`DK-2000 Frederiksberg (DK).
`
`(81) Designated States: AE, AL, AM, AT, AU, AZ, BA, BB, BG,
`BR, BY, CA, CH, CN, CR, CU, CZ, DE, DK, DM, DZ, EE,
`ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP,
`KE, KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA,
`MD, MG, MK, MN, MW, MX, NO, NZ, PL, PT, RO, RU,
`SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG,
`UZ, VN, YU, ZA, ZW, ARIPO patent (GH, GM, KE, LS,
`MW, SD, SL, SZ, TZ, UG, ZW), Eurasian patent (AM, AZ,
`BY, KG, KZ, MD, RU, TJ, TM), European patent (AT, BE,
`CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC,
`NL, PT, SE), OAPI patent (BF, BJ, CF, CG, CI, CM, GA,
`GN, GW, ML, MR, NE, SN, TD, TG).
`
`Published
`With international search report.
`Before the expiration of the time limit for amending the
`claims and to be republished in the event of the receipt of
`amendments.
`
`(54) Title: NEW SALT OF (2R,3R,4R)-3,4~DIHYDROXY-2-HYDROXYMETHYLPYRROLIDINE
`
`(57) Abstract
`
`This invention relates to (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpynolidine D-tartrate, preparation thereof and use as therapeutic
`
`agent.
`
`Apotex Exhibit 1017.001
`
`

`

`FOR THE PURPOSES OF INFORMATION ONLY
`
`Codes used to identify States party to the PCT on the front pages of pamphlets publishing international applications under the PCT.
`
`AL
`AM
`AT
`AU
`AZ
`BA
`BB
`BE
`BF
`BG
`BJ
`BR
`BY
`CA
`CF
`CG
`CH
`CI
`CM
`CN
`CU
`CZ
`DE
`DK
`EE
`
`Albania
`Armenia
`Austria
`Australia
`Azerbaijan
`Bosnia and Herzegovina
`Barbados
`Belgium
`Burkina Faso
`Bulgaria
`Benin
`Brazil
`Belarus
`Canada
`Central African Republic
`Congo
`Switzerland
`Cbte d’Ivoire
`Cameroon
`China
`Cuba
`Czech Republic
`Germany
`Denmark
`Estonia
`
`ES
`FI
`FR
`GA
`GB
`GE
`GH
`GN
`GR
`HU
`IE
`IL
`IS
`IT
`JP
`KE
`KG
`KP
`
`KR
`KZ
`LC
`LI
`LK
`LR
`
`Spain
`Finland
`France
`Gabon
`United Kingdom
`Georgia
`Ghana
`Guinea
`Greece
`Hungary
`Ireland
`Israel
`Iceland
`Italy
`Japan
`Kenya
`Kyrgyzstan
`Democratic People’s
`Republic of Korea
`Republic of Korea
`Kazakstan
`Saint Lucia
`Liechtenstein
`Sri Lanka
`Liberia
`
`LS
`LT
`LU
`LV
`MC
`MD
`MG
`MK
`
`ML
`MN
`MR
`MW
`MX
`NE
`NL
`NO
`NZ
`PL
`PT
`RO
`RU
`SD
`SE
`SG
`
`Lesotho
`Lithuania
`Luxembourg
`Latvia
`Monaco
`Republic of Moldova
`Madagascar
`The former Yugoslav
`Republic of Macedonia
`Mali
`Mongolia
`Mauritania
`Malawi
`Mex ico
`Niger
`Netherlands
`Norway
`New Zealand
`Poland
`Portugal
`Romania
`Russian Federation
`Sudan
`Sweden
`Singapore
`
`SI
`SK
`SN
`sz
`TD
`TG
`TJ
`TM
`TR
`TT
`UA
`UG
`US
`uz
`VN
`YU
`ZW
`
`Slovenia
`Slovakia
`Senegal
`Swaziland
`Chad
`Togo
`Tajikistan
`Turkmenistan
`Turkey
`Trinidad and Tobago
`Ukraine
`Uganda
`United States of America
`Uzbekistan
`Viet Nam
`Yugoslavia
`Zimbabwe
`
`Apotex Exhibit 1017.002
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`WO 00/55131
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`TITLE
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`1
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`PCT/DK00/00104
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`New salt of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine
`
`FIELD OF THE INVENTION
`
`This invention relates to (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine D-tartrate, its
`preparation and use as therapeutic agent.
`
`BACKGROUND OF THE INVENTION
`
`Diabetes
`Diabetes is characterised by an impaired glucose metabolism manifesting itself among other
`things by an elevated blood glucose level in the diabetic patients. Underlying defects lead to
`a classification of diabetes into two major groups: type 1 diabetes, or insulin demanding dia­
`betes mellitus (IDDM), which arises when patients lack β-cells producing insulin in their pan­
`creatic glands, and type 2 diabetes, or non-insulin dependent diabetes mellitus (NIDDM),
`which occurs in patients with an impaired β-cell function besides a range of other abnormali­
`ties.
`
`Type 1 diabetic patients are currently treated with insulin, while the majority of type 2 diabetic
`patients are treated either with sulfonylureas that stimulate β-cell function or with agents that
`enhance the tissue sensitivity of the patients towards insulin or with insulin. Among the
`agents applied to enhance tissue sensitivity towards insulin metformin is a representative
`example.
`
`In normal as well as in diabetics, the liver produces glucose in order to avoid hypoglycaemia.
`This glucose production is derived either from the release of glucose from glycogen stores or
`from gluconeogenesis, which is a de novo intracellular synthesis of glucose. In type 2 diabe­
`tes, however, the regulation of hepatic glucose output is poorly controlled and is increased,
`and may be doubled after an overnight fast. Moreover, in these patients there exists a strong
`correlation between the increased fasting plasma glucose levels and the rate of hepatic glu­
`cose production (reviewed in R.A. De Fronzo: Diabetes 37 (1988), 667 - 687; A. Consoli:
`
`Apotex Exhibit 1017.003
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`2
`Diabetes Care 15 (1992), 430 - 441; and J.E. Gerich: Horm.Metab.Res. 26 (1992), 18 -21).
`Similarly, hepatic glucose production will be increased in type 1 diabetes, if the disease is
`not properly controlled by insulin treatment.
`
`Since the liver in diabetes is known to have an increased glucose production, compounds
`inhibiting this activity are highly desirable. Recently, patent applications on inhibitors of the
`liver specific enzyme, glucose-6-phosphatase, which is necessary for the release of glucose
`from the liver, have been filed, for example German Offenlequngsschrift Nos. 4,202,183 and
`4,202,184 and Japanese patent application No. 4-58565. All these known compounds are
`benzene derivatives.
`
`Glycogen phosphorylase is another enzyme, which is necessary for the release of glucose
`from the liver. Substituted N-(indole-2-carbonyl)-glycinamides acting as glycogen phosphory­
`lase inhibitors are disclosed in PCT-publications No. WO96/39384 and WO96/39385 and in
`EP-A-0 846 464. Piperidine and pyrrolidine compounds acting as glycogen phosphorylase
`inhibitor are disclosed in PCT-publication No. WO95/24391, WO 97/09040, WO 98/40353
`and WO 98/50359.
`A compound which effectively can be used for treatment or preventing of diabetes is
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine.
`
`Obesity or appetite regulation
`Another field for the invention is obesity or appetite regulation.
`Obesity is a well-known risk factor for the development of many very common diseases such
`as atherosclerosis, hypertension and diabetes. The incidence of obese people and thereby
`also these diseases is increasing throughout the entire industrialised world.
`Due to its indirect but important effect as a risk factor in mortal and common diseases it will
`be important to find treatment for obesity or appetite regulation.
`
`Exercise, diet modification and food restriction will reduce body weight but for most patients,
`this is not feasible. Pharmacological treatment available up to date only consists of Sibutra­
`mine (acting via serotonergic mechanisms, Knoll Pharm) and Orlistat (reducing fat uptake
`from the gut, Roche Pharm) neither reducing body weight effectively nor acceptably.
`The term obesity implies an excess of adipose tissue. In this context obesity is best viewed
`as any degree of excess adiposity that imparts a health risk. The cut off between normal and
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`Apotex Exhibit 1017.004
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`3
`obese individuals can only be approximated, but the health risk imparted by the obesity is
`probably a continuum with increasing adiposity. The Framingham study demonstrated that a
`20% excess over desirable weight clearly imparted a health risk. (Mann GV N.Engl.J.Med
`291:226, 1974). In the United States a National Institutes of Health consensus panel on obe­
`sity agreed that a 20% increase in relative weight or a body mass index (BMI = body weight
`in kilograms divided by the square of the height in meters) above the 85th percentile for
`young adults constitutes a health risk. By the use of these criteria 20 to 30 percent of adult
`men and 30 to 40 percent of adult women in the United States are obese. (NIH, Ann Intern
`Med 103:147, 1985).
`Indeed, the prevalence of obesity has increased with 100% in most western countries the
`last 20 years, and this is very serious because even mild obesity increases the risk for pre­
`mature death, type 2 diabetes, coronary heart disease, hypertension, atherosclerosis, sleep
`apnea and respiratory problems, osteoarthritis, gallbladder disease and certain types of can­
`cer (endometrial, breast, prostate and colon). Because of the high prevalence of obesity and
`its health consequences, its prevention and treatment should be a high public health priority.
`
`When energy intake exceeds expenditure, the excess calories are stored in adipose tissue,
`and if this net positive balance is prolonged, obesity results, i.e. there are two components to
`weight balance, and an abnormality on either side (intake or expenditure) can lead to obe­
`sity.
`
`The regulation of feeding behaviour is incompletely understood. Certain is that brain neuro­
`chemicals located in specific hypothalamic nuclei regulate onset and termination of feeding.
`Several regulatory processes may influence these hypothalamic centres: Metabolic signals
`such as postprandial increases in plasma glucose and insulin; meal-induced gastric disten­
`sion is another possible inhibitory factor. Local control by brain neurochemicals and cate-
`cholamines/beta3-adrenoceptors (inhibits feeding and stimulates energy expenditure). Psy­
`chological, social, and genetic factors also influence food intake.
`
`At present a variety of techniques are available to effect initial weight loss. Unfortunately,
`initial weight loss is not an optimal therapeutic goal. Rather, the problem is that most obese
`patient eventually regain their weight. An effective means to establish and/or sustain weight
`loss is the major challenge in the treatment of obesity today.
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`Apotex Exhibit 1017.005
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`4
`Thus there remains today a need in the art for compositions and methods that are useful for
`the treatment or prophylaxis of obesity or appetite regulation.
`
`One object of the present invention is to provide compounds, which can effectively be used
`for the treatment or prophylaxis of obesity or appetite regulation.
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`A compound which effectively can be used for the treatment or prophylaxis of obesity or ap­
`petite regulation is (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine.
`
`(2R,3R,4R)-3,4’Dihydroxy-2-hydroxymethvlpyrrolidine
`(2R,3R,4R)-3,4-Dihydroxy-2-hydroxymethylpyrrolidine is a base with a pKa 8.22. As the
`base is an oil ordinary tablet formulation cannot be applied. Therefere there is a need to find
`a crystalline salt with suitable solid-state characteristics like a high melting point and low
`hygroscopicity.
`
`(2R,3R,4R)-3,4-Dihydroxy-2-hydroxymethylpyrrolidine, hydrochloride, was disclosed in
`example 2 of WO 97/09040. The compound was prepared according to the method
`described by Overkleeft et al., Tetrahedron 50 (1994), 4215-4224, which is incorporated
`herein by reference.
`
`However, the hydrochloride salt has some pharmaceutically undesirable properties. The
`hydrochloride salt is highly hygroscopic at relative room humidities above 50%
`and has a melting point of 116 °C, which is rather low. For the choice of tablet formulation
`process it would be a big advantage to have a salt with a higher melting point.
`
`For commercial use it is important to have a physiologically acceptable salt with good stability,
`good solubility, non-hygroscopicity, good bioavailability, good handling properties, like high
`melting point and a reproducible crystalline form.
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`
`SUMMARY OF THE INVENTION
`
`It has now been discovered that (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine, D-
`tartrate surprisingly has the above described desired properties.
`
`Apotex Exhibit 1017.006
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`WO 00/55131
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`5
`Within one aspect, the present invention provides (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-
`pyrrolidine,D-tartrate.
`
`Within another aspect of the invention there is provided a salt for the use as a medicament.
`
`Within another aspect of the invention there is provided a use of the salt for the preparation of
`a pharmaceutical composition for the treatment or preventing of diabetes.
`
`Within another aspect of the invention there is provided a use of a salt for the preparation of a
`pharmaceutical composition for the treatment or prophylaxis of obesity or appetite regulation.
`
`Within another aspect of the invention there is provided a pharmaceutical composition
`comprising (2R,3R,4R)-3,4-dihydroxy-2-hydroxy-methylpyrrolidine,D-tartrate optionally
`together with a pharmaceutically acceptable carrier or diluent.
`
`Within another aspect-of the invention there is provided a process for the preparation of
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine,D-tartrate which process comprises
`dissolving D-tartaric acid and (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine in a
`suitable solvent, and crystallizing the resulting salt from the solution.
`
`Within another aspect of the invention there is provided a process for the preparation of
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine, D-tartrate which process comprises
`dissolving D-tartaric acid and a salt of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine
`in a suitable solvent, and crystallizing the resulting salt from the solution.
`
`Furthermore a process for the preparation of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-
`pyrrolidine, D-tartrate is produced by reducing 2,3,5-Tri-O-benzyl-1,4-dideoxy-1,4-imino-D-
`arabinitol with Pd/C in glacial acetic acid. The formed, oily (2R,3R,4R)-3,4-dihydroxy-2-
`hydroxymethyl-pyrrolidine,acetate is dissolved in methanol and added to a solution
`containing D-tartraric acid at reflux and (2R,3R,4R)-3,4-dihydroxy-2-
`hydroxymethylpyrrolidine, D-tartrate is formed and can be isolated by filtration. Ion exchange
`chromatography is thereby avoided.
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`Apotex Exhibit 1017.007
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`6
`Within another aspect of the invention there is provided a method of using the salt according to
`the invention for the treatment or preventing of diabetes and a method of using the salt
`according to the invention for the treatment or prophylaxis of obesity or appetite regulation.
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`DETAILED DESCRIPTION OF THE INVENTION
`
`The present invention relates to the D-tartrate salt of (2R,3R,4R)-3,4-dihydroxy-2-
`hydroxymethylpyrrolidine having the structural formula I as shown herein below.
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`Hereinafter the above compound of formula I is referred to as compound I.
`
`A salt of compound I is provided in the form of crystals, which have good stability
`characteristics in e.g. non-hygroscopicity, good solubility in e.g. water, good bioavailability,
`good handling properties in e.g. high melting point, and a reproducible crystalline form.
`
`The present invention also provides a process for the preparation of a salt of compound I,
`which process comprises dissolving compound I in a suitable solvent, and dissolving a specific
`acid, in the same kind of solvent, and adding the solution of the acid to the solution of
`compound I, and crystallizing the resulting salt from the solution. Examples of the common
`solvents include but are not limited to organic solvents in particular lower aliphatic alcohol’s
`such as methanol, ethanol, 2-propanol, 2-butanol, 1-hexanol and solvents like acetone,
`isobutylmethylketone and tetrahydrofuran. Preferred solvents are methanol, ethanol, 2-
`propanol and acetone. The mixture of the components is conveniently performed at
`temperatures from 40° C to reflux before cooling down to 0-5°C and collection of the crystals
`by filtration. The speed of cooling down, the crystallisation temperature and the solvent may
`have influence on the crystalline form obtained. Optionally, seeding crystals are added if
`precipitation has not occurred within 1-2 hours after mixing.
`
`Apotex Exhibit 1017.008
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`7
`
`The present invention also provides a pharmaceutical composition comprising a salt of
`compound I optionally together with a pharmaceutically acceptable carrier or diluent.
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`A salt of compound I may be used in human and veterinary medicine for the treatment or
`preventing of diabetes and for the treatment or prophylaxis of obesity or appetite regulation.
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`A salt of compound I may be used in human and veterinary medicine for the treatment or
`preventing of diabetes, and especially non-insulin dependent diabetes (NIDDM or type 2
`diabetes) including overnight or meal treatment.
`
`A still further objection of this invention is to provide a salt of compound I which can
`effectively be used as an inhibitor of glucose production from the liver.
`
`A still further objection of this invention is to provide a salt of compound I which can
`effectively be used as glycogen phosphorylase inhibitor.
`For use within the present invention (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine,D-
`tartrate may be formulated with a pharmaceutically acceptable carrier or excipient to provide a
`medicament for parenteral, oral, nasal, rectal, pulmonal, buccal, subdermal or intradermal or
`transdermal administration according to conventional methods. Formulations may further
`include one or more diluents, fillers, emulsifiers, preservatives, buffers, excipients, etc. and
`may be provided in such forms as liquids, powders, emulsions, suppositories, liposomes,
`transdermal patches, controlled release, dermal implants, tablets, etc.
`
`One skilled in this art may formulate the compound in an appropriate manner, and in accor­
`dance with accepted practices, such as those disclosed in Remington's Pharmaceutical Sci­
`ences, 1985 or Remington's Pharmaceutical Sciences, Gennaro, ed., Mack Publishing Co.,
`Easton, PA, 1990, or Remington: The Science and Practice of Pharmacy, 19th Edition (1995),
`which are incorporated by reference. The compositions may appear in conventional forms, for
`example capsules, tablets, aerosols, solutions, suspensions or topical applications.
`
`The pharmaceutical carrier or diluent employed may be a conventional solid or liquid carrier.
`Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pec­
`tin, acacia, magnesium stearate, stearic acid or lower alkyl ethers of cellulose. Examples of
`
`Apotex Exhibit 1017.009
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`8
`liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, poly­
`oxyethylene or water.
`
`Similarly, the carrier or diluent may include any sustained release material known in the art,
`such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.
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`The compositions of this invention are usually adapted for oral administration.
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`For oral administration (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine,D-tartrate is
`prepared in a suitable form, such as a tablet or capsule. Typically, the salt of compound I is
`combined with a carrier and molded into a tablet. Suitable carriers in this regard include starch,
`sugars, dicalcium phosphate, calcium stearate, magnesium stearate and the like. Such
`compositions may further include one or more auxiliary substances, such as wetting agents,
`emulsifiers, preservatives, stabilizers, colouring additives, etc.
`
`Pharmaceutical compositions are administered one or more times per day or week. An
`effective amount of such a pharmaceutical composition is the amount that provides a clinically
`significant effect. Such amounts will depend, in part, on the particular condition to be treated,
`age, weight, and general health ofthe patient, and other factors evident to those skilled in the
`art.
`
`The pharmaceutical compositions may be administered in unit dosage form one or more times
`per day or week. In the alternative, they may be provided as controlled release formulations
`suitable for dermal implantation. Implants are formulated to provide release of active
`compound over the desired period of time, which can be up to several years. Controlled-
`release formulations are disclosed by, for example, Sanders et al., J Pharm Sci 73 (1964),
`1294 - 1297,1984; U.S. Pat. No. 4,489,056; and U.S. Pat. No. 4,210,644, which are
`incorporated herein by reference.
`
`The composition is usually presented as a unit dose composition containing 0.1 -1000 mg of a
`salt of compound I in accordance with the invention for oral dosing. Typical dosage for diabetes
`effect would vary between 0.1- 750 mg, preferably between 1 - 500 mg per day either once or
`divided in 2 or 3 doses when administered orally or 2 or 3 times per week or once weekly or
`once per 14 days.
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`Apotex Exhibit 1017.010
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`
`Preferred unit dosage forms include in solid form, tablets or capsules, in liquid form, solutions,
`suspensions, emulsions, elixirs or capsules filled with the same, or in form of sterile injectable
`solutions, or patches, vagitories, vaginal rings or long lasting implantates.
`
`The composition of this invention may be formulated by conventional methods of galenic
`pharmacy.
`
`Conventional excipients are such pharmaceutically acceptable organic or inorganic carrier
`substances suitable for parenteral or oral application which do not deleteriously react with the
`active compound I.
`
`Examples of such carriers are water, salt solutions, alcohols, polyethylene glycols,
`polyhydroxyethoxylated castor oil, syrup, peanut oil, olive oil, gelatin, lactose, terra alba,
`sucrose, agar, pectin, acacia, amylose, magnesium stearate, talc, silicic acid, stearic acid, fatty
`acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, hydroxymethylcellulose
`and polyvinylpyrrolidone and calcium phosphates.
`
`The pharmaceutical preparations can be sterilized and mixed, if desired, with auxiliary agents,
`such as binders, lubricants, preservatives, disintegrants, stabilizers, wetting agents,
`emulsifiers, salt for influencing osmotic pressure, buffers and/or colouring substances and the
`like, which do not deleteriously react with the active compound.
`
`Particularly suitable for parenteral application are injectable solutions or suspensions,
`preferably aqueous solutions with the active compound dissolved in polyhydroxylated castor
`oil.
`
`Particularly suitable for oral administration are tablets, dragees, or capsules having talc and/or
`a carbohydrate carrier or binder or the like, the carrier preferably being lactose or calcium
`phosphate and/or corn starch and/or potato starch. A syrup, elixir or like can be used when a
`sweetened vehicle can be employed.
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`Apotex Exhibit 1017.011
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`A typical tablet, which may be prepared by conventional tabletting techniques, contains: 50 mg
`of compound I in form of the salt, 100 mg of lactose, 30 mg of corn starch, 3 mg of talc powder,
`3 mg of colloidal silicon dioxide and 2 mg of magnesium stearate.
`
`The present invention is further illustrated by the following examples, which, however, are not
`to be construed as limiting the scope of protection. The features disclosed in the foregoing
`description and in the following examples may, both separately and in any combination thereof,
`be material for realising the invention in diverse forms thereof.
`
`(2R,3R,4R)-3,4-Dihydroxy-2-hydroxymethylpyrrolidine, D-tartrate is synthesized, purified and
`crystallized as described in the following examples.
`
`Examples
`
`Compound I, (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine, has the structural for­
`mula I as shown herein below.
`
`HO OHV /
`\ ZOH
`
`NH
`
`Formula I
`
`Example 1
`The hydrochloride salt of compound I is disclosed in example 2 of WO 97/09040. The hydro­
`chloride salt of compound I may be prepared as described by Overkleeft et al., Tetrahedron
`50 (1994), 4215-4224, which is incorporated herein by reference.
`
`In brief, the hydrochloride salt of compound I can be prepared as follows:
`2,3,5-Tri-O-benzyl-D-arabinofuranose is oxidised via DMSO/acetic acid anhydride to its lac­
`tone. 2,3,5-Tri-O-benzyl-D-arabino-1,4-lactone was reacted with ammonia to furnish the ara­
`binoamide which then is oxidised via Dess Martin agent to the 4-oxo-arabinoamide. Treat­
`ment with ammonia and affords a 1:1 C4 isomeric mixture of the hydroxylactams. The iso­
`meric mixture is reduced with sodium cyanoborohydride to afford pure 2,3,5-Tri-O-benzyl-D-
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`arabino-1,4-lactam after purification. The lactame is reduced to 2,3,5-Tri-O-benzyl-1,4-
`dideoxy-1,4-imino-D-arabinitol with LiAIH4. Subsequent reduction with Pd/C containing HCI
`followed by purification and crystallisation led to pure (2R,3R,4R)-3,4-dihydroxy-2-
`hydroxymethylpyrrolidine.HCl.
`
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine.
`A normal glass column for flash chromatography (diameter 2 cm) was filled with ion ex­
`change resin from Rohm & Haas Ambersep 900 OH to a height of 10.4 cm. 50 ml water was
`added. The column was rinsed with 50 ml water and then with 150 ml 1M NaOH over a pe­
`riod of 30 min. then, the column was washed with 3x50ml water and 3x50ml methanol. 2.00
`g compound I, HCI was dissolved in 20 ml methanol and run through the column 5 times
`over a total period of 10 min. The final methanolic solution was collected and the solvent re­
`moved. The residual brownish oil was stripped with ethanol to removed small amounts of
`water. The final yield of compound I free base was 1.66 g (93%) and contained less than
`0.1% ionogen chlorine measured by elemental analysis.
`
`(2R,3R,4R)-3,4-Dihydroxy-2-hydroxymethylpyrrolidine, D-tartrate.
`A solution of D-tartaric acid (225 mg, 1.5 mmol) in methanol (1 ml) is added to a solution of
`compound I (200 mg, 1.5 mmol) in methanol (4 ml). The mixture is cooled at -18 °C for 2
`hours. The formed precipitate is collected, washed with cold methanol (1ml) and dried to give
`white crystals of (2R,3R,4R)-3,4-dihydroxy-2-hydroxy methyl-pyrrolidine, D-tartrate (Yield:
`250 mg) (CsH^f^-C^Og; MW: 283,24g/mol). Melting point of 148 °C.
`An evaluation of the hygroscopicity was carried out by exposing the salt to increasing humid­
`ity and measuring the mass increase in a Dynamic Vapour Sorption Instrument (DVS). The
`critical relative humidity value is found to be above 80%.
`
`Example 2
`A more direct way to synthesise 3,4-dihydroxy-2-hydroxymethylpyrrolidine, D-tartrate is by
`reduring 2,3,5-Tri-O-benzyl-1,4-dideoxy-1,4-imino-D-arabinitol with Pd/C in glacial acetic
`acid. The formed, oily (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine,acetate is dis­
`solved in methanol and added to a solution containing D-tartraric acid at reflux and
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine, D-tartrate is formed and can be iso­
`lated by filtration. Ion exchange chromatography is thereby avoided.
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`Synthesis of (2R,3R,4R)-3,4-dihvdroxy-2-hydroxymethylpyrrolidine,D-tartrate via
`(2R,3R,4R)-3,4-dihvdroxv-2-hvdroxymethvlpvrrolidine, acetate
`10g (25mmol) of 2,3,5-Tri-O-benzyl-1,4-dideoxy-1,4-imino-D-arabinitol is dissolved in 40ml
`glacial acetic acid. Heat is developed and the solution gets dark. 1.2g of 10% Pd/C (50%
`wet) is added and stirred overnight at 20-25°C under nitrogen atmosphere. The catalyst is
`removed by filtration and washed with 10ml of glacial acetic acid. Another 1.2g of 10% Pd/C
`(50% wet) is added and the mixture is left under hydrogen atmosphere at 3bar and 50°C.
`Full conversion is obtained after 41/2h. The mixture is cooled to 20-25°C and filtered. The fil­
`trate is evaporated and stripped 3 times with 20ml of a 50:50 methanol/toluene mixture to
`remove excess acetic acid leaving 7.0g (>100%) of an oily, dark residue with an extensive
`odour of acetic acid.
`
`The oily residue is dissolved in 15ml methanol and slowly added to 15ml refluxing methanol
`containing 4.4g (29 mmol) D-tartaric acid. When around one third of the mixture is added
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine,D-tartrate begins to crystallise out of
`the solution. The temperature is kept at reflux for additional 1/2h after all (2R,3R,4R)-3,4-
`dihydroxy-2-hydroxymethylpyrrolidine,acetate is added and then cooled to 20-25°C by stand­
`ing overnight. The next day the slurry is filtered and the filter cake dried to constant weight
`leaving 6.1g (86%) of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine, D-tartrate
`(CsHuNCtyCiHgOg; MW: 283,24g/mol) as off white crystals. Melting point of 147°C.
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`Apotex Exhibit 1017.014
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`WO 00/55131
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`CLAIMS
`
`13
`
`PCT/DK00/00104
`
`1. (2R,3R,4R)-3,4-Dihydroxy-2-hydroxymethylpyrrolidine, D-tartrate.
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`2. A process for the preparation of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine, D-
`tartrate according to claim 1, which process comprises dissolving D-tartaric acid and
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine in a suitable solvent, and crystallizing
`the resulting salt from the solution.
`
`3. A process for the preparation of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine, D-
`tartrate according to claim 1, which process comprises dissolving D-tartaric acid and a salt of
`(2R,3R,4R)-3,4-dihydroxy-2-hydroxymethylpyrrolidine in a suitable solvent, and crystallizing
`the resulting salt from the solution.
`
`4. A process for the preparation of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine, D-
`tartrate according to claim 3, wherein the salt of (2R,3R,4R)-3,4-dihydroxy-2-
`hydroxymethylpyrrolidine is acetate.
`
`5. A process for the preparation of (2R,3R,4R)-3,4-dihydroxy-2-hydroxymethyl-pyrrolidine, D-
`tartrate according to anyone of the claims 2-4, wherein the solvent is methanol.
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`6. A pharmaceutical composition comprising (2R,3R,4R)-3,4-dihydroxy-2-hydroxy-
`methylpyrrolidine, D-tartrate according claim 1 optionally together with a pharmaceutically
`acceptable carrier or diluent.
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`7. The pharmaceutical composition according to claim 6 in the form of a dosage unit containing
`about 0.1-1000 mg per day.
`
`8. A pharmaceutical composition for the treatment or preventing of diabetes in a patient
`comprising a therapeutically effective amount of a salt according to claim 1, together with a
`pharmaceutically acceptable carrier or diluent.
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`Apotex Exhibit 1017.015
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`14
`9. A pharmaceutical composition for the treatment or prophylaxis of obesity or appetite
`regulation in a patient comprising a therapeutically effective amount of a salt according to claim
`1, together with a pharmaceutically acceptable carrier or diluent.
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`10. A salt according to claim 1 for use as a medicament.
`
`11. Use of a salt according to claim 1 for the preparation of a pharmaceutical composition for
`the treatment or preventing of diabetes.
`
`12. Use of a salt according to claim 1 for the preparation of a pharmaceutical composition for
`the treatment or prophylaxis of obesity or appetite regulation.
`
`13. A method for the treatment or preventing of diabetes comprising administering to a patient
`an effective amount of a salt according to claim 1.
`
`14. A method for the treatment or prophylaxis of obesity or appetite regulation comprising
`administering to a patient an effective amount of a salt according to claim 1.
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`Apotex Exhibit 1017.016
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`INTERNATIONAL SEARCH REPORT
`
`International application No.
`PCT/DK 00/00104
`
`A. CLASSIFICATION OF SUBJECT MATTER
`
`IPC7: C07D 207/12, A61K 31/40
`According to International Patent Classification (IPC) or to both national classification and IPC
`B. FIELDS SEARCHED
`.Minimum documentation searched (classification system followed by classification symbols)
`
`IPC7: C07D
`Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched
`
`SE,DK,FI,N0 classes as above
`
`Electronic data base consulted during the international search (name of data base and, where practicable, search terms used)
`
`C. DOCUMENTS CONSIDERED TO BE RELEVANT
`
`Category* Citation of document, with indication, where appropriate, of the relevant passages
`
`Relevant to claim No.
`
`A
`
`WO 9709040 Al (NOVO NORDISK A/S), 13 Marc