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Title:
ARACHIDONIC ACID ANALOGS AND METHODS FOR ANALGESIC TREATMENT USING SAME
Document Type and Number:
WIPO Patent Application WO/2011/066414
Kind Code:
A1
Abstract:
The present invention provides arachidonic acid (AA) analogs and compositions containing those analogs as active agents for use in analgesic treatments. Various methods of r manufacturing the inventive compounds are provided and pharmaceutical formulations, including injectable and oral dosages, are described. The analogs are additionally useful as antipyretic compositions and in related fever reducing treatments.

Inventors:
BROSTROM LANE (US)
FALCK JOHN R (US)
Application Number:
US2010/058041
Publication Date:
June 03, 2011
Filing Date:
November 24, 2010
Export Citation:
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Assignee:
CYTOMETIX INC (US)
BROSTROM LANE (US)
FALCK JOHN R (US)
International Classes:
C07C233/09; A61K31/16; A61K31/164; A61K31/17; A61P29/00; C07C237/22; C07C275/20
Domestic Patent References:
WO1994012466A11994-06-09
Foreign References:
US4256108A1981-03-17
US0160452A1875-03-02
US4265874A1981-05-05
US4663308A1987-05-05
GB8900581W1989-05-25
US4777049A1988-10-11
Other References:
HARDY ET AL., ALIMENT. PHARMACOL. THERAP., vol. 1, 1987, pages 273 - 280
FRIEND ET AL., J. MED. CHEM., vol. 27, 1984, pages 261 - 2681
REAGAN-SHAW, FASEB J., vol. 22, 2007, pages 659 - 651
Attorney, Agent or Firm:
LEECK, Charles, L. (411 E. Wisconsin Ave.Milwaukee, WI, US)
Download PDF:
Claims:
128833.00010 PCT Application

CLAIMS

What is clai med is:

1. A compound having the structure:

whe ¾in:

Ri is H, or a C] -C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl or C3-C6 cycl ^alkenyl which is unsubstituted or substituted with at least one hydroxyl groi p; and

R2 is H, or a C1-C3 alkyl; or

Ri and R2 form a C3-C6 heterocyclic ring with the nitrogen bonded to said Ri a nd R2;

R4 is H, or a Ci-Ce alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl or C3-C6 cycl Dalkenyl;

R5 is a C1-C6 alkyl, Ci-Ce alkoxy, or C2-C6 alkyl ether which is unsubstituted or substituted with one or more of hydroxyl, phenyl, phenyloxy, or fluo -ine, or R5 is NR7R8, or C(0)NR7R8 in which R7 and Rg are independently sele cted from H, a C1-C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl, or C3-C6 cycl oalkenyl group;

45

QB\ 1 1803388.

128833.00010 PCT Application

R6 is H, or a C C6 alkyl, C2-C6 alkenyl, C3-C6 cycloalkyl, or C3-C6 eye oalkeny! group; and

n is 0, 1 or 2; or

a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1 , wherein R3 is

3. The compound according to any one of claims 1 or 2, wherein R5 is a linear C5 alkyl group.

4. The compound according to any one of claims 1 -3, wherein R) is a eye opropyl group and R2 is H.

5. The compound according to any one of claims 1 -4, wherein R4 is a C3 i sopropyl group.

6. The compound according to any one of claims 1 -5, wherein n is i .

wherein the compound has

46

QB\1 1803388. 1

128833.00C 10 PCT Application

8. A compound having a structure selected from the group consisting

47

QB\1 1803388.

128833.00010 PCT Application

9. A composition comprising a compound according to any one of claims 1 -8 and a phanr aceutically acceptable carrier.

10. The composition of claim 9, wherein said composition is in the form of an injectable d> >sage.

1 1. The composition of claim 9, wherein said composition is in the form of an oral dosage,

12. The composition of claim 9, wherein said composition is in the form of an oil-in-water emulsion.

13. The composition of claim 9, wherein said composition is in the form of an anhydrous e mulsion.

14. The composition of claim 9, wherein said composition comprises a cyclodextrii .

15. The composition of claim 9, further comprising an anesthetic agent.

16. A kit for providing analgesia to a subject, comprising a compound according any one of claims 1 -8 and a delivery device to administer said compound to the subject.

17. A method of providing analgesia to a subject, comprising administering to a subject a t herapeutically effective amount of a compound according to any one of claims 1 -8, whereby analgesia is provided in said subject.

18. The method of claim 17, wherein administering said compound is by intravenous injection.

19. The method of claim 17, wherein administering said compound is by oral delivery.

20. The method of claim 17, wherein administering said compound is by bolus intravenous injection.

21 . The method of claim 17, wherein administering said compound is by continuous ntravenous infusion.

22. Use of a compound according to any one of claims 1 -8 for the manufacture of a medicament for providing analgesia in a subject.

23. A compound according to any one of claims 1 -8 for use in providing analgesia in a subject.

48

QB\1 1803388.

128833.00010 PCT Application

24. A method of providing an arachidonic acid (AA) analog, comprising the steps of :

A method of providing an arachidonic acid (AA) analog, comprising the

OH OH

I TsCI, Py

49

QB\1 1803388.1

128833.000 10 PCT Application

26. A method of reducing fever in a subject, comprising administering to a subject a the rapeutically effective amount of a compound according to any one of claims 1 -8, whereby fever is reduced in said subject.

27. Use of a compound according to any one of claims 1-8 for the manufacture of a medicament for reducing fever in a subject.

28. A compound according to any one of claims 1 -8 for use in reducing fever in a subject.

50

QB\1 1803388.1

Description:
128833.00310 PCT Application

ARACHIDONIC ACID ANALOGS AND METHODS FOR ANALGESIC TREATMENT USING SAME

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This International Application claims the benefit of U.S. Provisional applicatior 61 /264,434, filed November 25, 2009, which is incorporated by reference herein in il s entirety for all purposes.

STA TEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR

DEVELOPMENT

[0002] Not Applicable.

FIELD OF THE INVENTION

[0003] This invention relates to the treatment of pain. In particular this invention is directed to arachidonic acid (AA) analogs and their use in analgesic treatment.

BACKGROUND OF THE INVENTION

[0004] The pain pathway begins in the periphery with nociceptors that innervate skin, muscle, tendon or bone targets. Activated or sensitized nociceptors transmit noxious informatio n to the spinal dorsal horn where spinal neurons then transmit information to rostral cen :ers in the thalamus, reticular formation and midbrain. Other neurons carry the informatio si to the somatosensory cortex where pain is interpreted. Nociceptive informatio n transmitted through the spinal cord is heavily modulated by central neurons whose axo ns descend from the midbrain and other rostral areas to the spinal cord, and these descending pathways can be either inhibitory or facilitory.

[0005] Neurons contain a variety of voltage-gated ion channels. The voltage-gated K+ and Ni .+ channels regulate the excitability of neuronal cells and play a crucial role in setting the perceptual threshold of pain. The ability to modulate the activity of K+ or Na+ ion channs Is in neuronal cells is important for regulating the transmission of pain signals.

1

QBM 180338S . 1 128833.00010 PCT Application

[0006] Epoxyeicosatrienoic acids (EETs) are produced from arachidonic acid via cytochrome P450 (CYP) epoxygenases. EETs regulate inflammation, angiogenesis, cellular pro liferation, ion transport and steroidogenesis. In many issues, EET levels are regulated, i iter alia, through their metabolism to v c-diols (v c-dihydroxyeicosatrienoic acids; DiHl iTrE) via the enzyme soluble epoxide hydrolase (EPHX2).

[0007] While some types of pain are effectively managed with opioids such as morphine c r non-steroidal anti-inflammatory drugs (NSAIDS) such as aspirin or ibuprofen, opioids and NSAIDS both have numerous undesirable side effects. For instance, o| lioids frequently cause dependence and withdrawal problems in users. The use of opio ds in the management of pain is further limited due to impairment of Na+/ + -ATPase a tivity after opiate treatment, a possible mechanism of tolerance/addiction. Similarly, NSAIDS can cause hypertension, ulcer perforation, upper gastrointestinal bleeding ar d even death in severe cases.

[0008] Acetaminophen is one of the most widely used drugs in the world for treatment o f pain and fever; probably the most commonly prescribed medicine in children. Over 600 products contain acetaminophen including OTC pain, cold and flu remedies and prescription medications like Vicodin. It has a unique position among analgesic d -ugs. Unlike NSAIDs, it is considered an ineffective anti-inflammatory, but does not pnduce gastrointestinal damage or untoward cardio-renal effects; unlike opiates, it i ί ineffective in pain arising from smooth muscle spasm , but has no depressant effect on re spiration. The acetaminophen metabolite that produces analgesia is AM404 - now knowt to provide analgesia through CB 1 and TRPV1 receptors. Fig. 1 illustrates the metabo ism of acetaminophen to A 404 and, in addition, the less desirable molecule NAPQI. Unfortunately acetaminophen is toxic in high doses and is responsible for the majority of the acute liver failure cases in the United States. NAPQI is the molecule largely beli eved to be responsible for liver failure.

[0009] Accordingly, a need exists for improved analgesic treatment that avoids the above-men :ioned side effects but provides an effective and safe treatment for pain.

2

QB\ 1 1803388. 1 128833.00( 10 PCT Application

SUMMARY OF THE INVENTION

[0010] -lere, the inventors demonstrate novel compositions of arachidonic acids analogs anc methods of use thereof for treatment of pain. The invention is based, in part, on the understanding that arachidonic acid (AA) is a catalyzable substrate for cytochrome P450 epox) genase (CYP4X1) in neurons. AA is converted to four regioisomers of EETs (i.e., 5,6-EI T; 8,9-EET; 1 1 ,12-EET and 14,15-EET) by CYP4X1 , and application of nanomolar concentrations of EETs (e.g., 1 1 ,12-EET) induces suppression of the outward + current and inward Na+ current, effectively altering the cellular membrane potential and polarization in neurons. EETs (or other P450 epoxygenase-derived epoxides) and certain sele ;ted agonist analogs may therefore regulate neuronal function and contribute to the modi lation and treatment of pain.

[0011] Accordingly, the invention encompasses in a first aspect certain compounds that are AA analogs having the structure:

wherein: Ri is H, or a Ci-C 6 alkyl, C 2 -C 6 alkenyl, C 3 -C 6 cycloalkyl or C 3 -C 6 cycloalkenyl which is unsubstituted or substituted with at least one hydroxyl group; and R 2 is H, or i C1-C3 alkyl; or Ri and R 2 form a C 3 -C 6 heterocyclic ring with the nitrogen bonded to said Ri and R 2 ;

in which:

R4 is H, or a Ci-Ce alkyl, C 2 -C 6 alkenyl, C3-C6 cycloalkyl or C 3 -C 6

cycloalkenyl; R 5 is a C 1 -C6 alkyl, C 1-C6 alkoxy, or C 2 -C 6 alkyl ether which is unsubstitut :d or substituted with one or more of hydroxyl, phenyl, phenyloxy, or fluorine, or R 5 is NR 7 Rg, or C(0)NR 7 Rg in which R 7 and R 8 are independently selected from H, a Ci-C 6 alkyl , C 2 -C 6 alkenyl, C 3 -C 6 cycloalkyl, or C 3 -C 6 cycloalkenyl group; R6 is H, or a

3

QB\1 1803388. 1 128833.00( 10 PCT Application

Ci-C 6 alkyl, C 2 -C 6 alkenyl, C 3 -C 6 cycloalkyl, or C 3 -C 6 eycloalkenyl group; and n is 0, 1 or 2; or a pharmaceutically acceptable salt thereof.

[0012]

QB\I 1803388. 1 128833.00C 10 PCT Application

[0013] \ particularly preferred compound according to the invention has the structure:

[0014] Compounds according to the invention are, in certain embodiments, provided in the form of a composition comprising a compound as described and claimed herein in combinatio ι with a pharmaceutically acceptable carrier. Particularly preferred compositions are in the form of injectable dosages or oral dosages. Certain compositions of the inver tion may be provided in the form of oil-in-water emulsions, while other compositions may be in form of anhydrous emulsions or lyophilized preparations.

Compositic ns of the invention may, in certain delivery vehicle formulations, include a cyclodextri ι with the compound.

[0015] n another aspect, the invention encompasses a kit for providing analgesia to a subject. Su ch a kit includes a compound as described and claimed herein and a delivery device to administer the compound to the subject.

[0016] The present invention further provides methods of providing analgesic treatment it i a subject, particularly the reduction of pain in a subject. Such methods include steps of administering to a subject a therapeutically effective amount of a compound is described and claimed herein, whereby analgesia is provided to the subject. Administra ion may be performed by, e.g., intravenous injection in bolus or continuous infusion fas hion, or by oral dosing with a tablet or capsule.

[0017] n yet another embodiment, the invention encompasses the use of AA analogs according t> the invention for the manufacture of a medicament for providing analgesia in a subject As well, the present invention further contemplates compounds according to the inventic n for use in providing analgesia in a subject.

[0018] The invention further provides a method of reducing fever in a subject. Such a method ir eludes the step of administering to a subject a therapeutically effective

5

QB\1 1 803388. 128833.000 10 PCT Application amount of a compound described and claimed herein, whereby fever is reduced in the subject. Of course, the invention encompasses methods of using an inventive compound for the mam ifacture of a medicament for reducing fever in a subject.

[0019] I n another aspect, the invention provides a method of providing an AA analog includes ste >s of:

OH I OH

TsCI Py

thereby presiding an AA analog according to the invention.

[0020] Another method of providing an AA analog includes steps of:

6

QB\1 1803388. : 128833.000 , 0 PCT Application

[0021] Ί he presently-described and claimed compounds and methods provide various advantages »ver prior compounds and methods in that they provide for analgesic effect with a reduc tion in the side effects encountered with prior analgesics.

[0022] ( >ther objects, features and advantages of the present invention will become apparent aft :r review of the specification, claims and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0023] I igure 1 depicts a general schematic illustrating acetaminophen metabolism.

[0024] I igure 2 illustrates the chemical structures of CMX-020 and the

acetaminopl ien metabolite AM404.

[0025] I igure 3 provides exemplary AA analogs of the invention including the compound ( !MX-020.

[0026] I igure 4 provides dose response and time profile data for CMX-020 and morphine as measured in the Tail-flick Assay. At 100% MPE, graphs are separated to better distin juish.

[0027] Figure 5 depicts Writhing Assay data in terms of Bolus Dose Response and Infusion Co parison for CMX-020, Morphine, and Perfalgan. At 100% MPE, graphs are separated to better distinguish.

[0028] Figure 6 provides CMX-020 Plasma Concentration in mouse measured by LC/MS.

[0029] I 'igure 7 illustrates the antipyretic (i.e., fever reducing) effect of compound CMX-020 a 3 compared to morphine in mice.

DETAILED DESCRIPTION OF THE INVENTION

I. IN GENERAL

[0030] l iefore the present materials and methods are described, it is understood that this inventit n is not limited to the particular methodology, protocols, materials, and reagents de: cribed, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by any later-filed n ^provisional applications.

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QB\1 1803388.1 128833.00010 PCT Application

[0031] 11 must be noted that as used herein and in the appended claims, the singular forms "a", " an", and "the" include plural reference unless the context clearly dictates otherwise. . \s well, the terms "a" (or "an"), "one or more" and "at least one" can be used interchange ibly herein. It is also to be noted that the terms "comprising", "including", and "having " can be used interchangeably.

[0032] Unless defined otherwise, all technical and scientific terms used herein have the same mi anings as commonly understood by one of ordinary skill in the art to which this inventk n belongs. Although any methods and materials similar or equivalent to those descri sed herein can be used in the practice or testing of the present invention, the preferred m ;thods and materials are now described. All publications and patents specifically mentioned herein are incorporated by reference for all purposes including describing e nd disclosing the chemicals, instruments, statistical analysis and

methodolog ies which are reported in the publications which might be used in connection with the inv sntion. All references cited in this specification are to be taken as indicative of the level _>f skill in the art. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

II. THI . INVENTION

[0033] . ^rachidonic acid (AA) analogs of the invention have analgesic effects similar to morphin< and other opioid analgesics. However, these compounds have a different mechanism of action than opioid analgesics. The inventors' preliminary tests have shown that AA am logs do not have the addiction side effects common in many conventional pain treatm ints. As well, preliminary results show that a number of different delivery options are possible, including brain injections, DRG injections, intraperitoneal injections, i ntranasal administration, blood injections, transdermal, or oral delivery. Chemical a lalogs of AA may be engineered or particularly delivered to have a more sustained e: Fect than traditional analgesics. Liposomes, mycelles, cyclodextrins, and emulsifiers can be used in to make AA analog preparations more soluble and easier to administer md/or more stable. A particularly preferred AA analog described herein is designated I1MX-020, which is chemically similar to the 14,15-EET (epoxyeicosatrienoic acid). The 14, 15-EET was observed to be the most potent analgesic of the natural EETs.

8

QB\ 1 1803388. 128833.000 10 PCT Application

However, C VlX-020 is a much more potent and longer lasting analgesic than the natural 14, 15-EET. The present invention further encompasses chemical variations based on CMX-020, i vhich are envisioned to be useful in analgesic treatment (exemplary such compounds ire illustrated in Fig. 3). The present compounds, including CMX-020, have also demons trated fever reducing effects and are further envisioned to find use as antipyretic c ompositions and in fever reducing therapies.

[0034] i LS used herein, "subject" means mammals and non-mammals. "Mammals" means any r iember of the class Mammalia including, but not limited to, humans, non- human prim ates such as chimpanzees and other apes and monkey species; farm animals such as cattl 2, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; lat oratory animals including rodents, such as rats, mice, and guinea pigs; and the like. Examples of non-mammals include, but are not limited to, birds, and the like. The term "s ibject" does not denote a particular age or sex.

[0035] i \s> used herein, "administering" or "administration" includes any means for introducing a compound of the present invention into the body, preferably into the systemic cir culation. Examples include but are not limited to oral, nasal, otic, ophthalmic, buccal, subl ngual, pulmonary, transdermal, transmucosal, as well as subcutaneous, intraperiton :al, intravenous, epidural and intramuscular injection.

[0036] i . "therapeutically effective amount" means an amount of a compound that, when admir istered to a subject for treating a disorder, condition, or disease, is sufficient to effect suc h treatment for the disorder, or condition, or disease. The "therapeutically effective an ount" will vary depending on the compound, the disorder, or condition, or disease stati : being treated, the severity or the disorder, or condition, or disease treated, the age and relative health of the subject, the route and form of administration, the judgment of the attending medical or veterinary practitioner, and other factors.

[0037] 1 'or purposes of the present invention, "treating" or "treatment" describes the managemer t and care of a patient for the purpose of combating the disease, condition, or disorder. Tl e terms embrace both preventative, i.e., prophylactic, and palliative treatments. Treating includes the administration of a compound of present invention to prevent the anset of the symptoms or complications, alleviating the symptoms or complicatio ns, or eliminating the disease, condition, or disorder.

9

QB\1 1803388. 128833.000 10 PCT Application

[0038] compound is administered to a patient in a therapeutically effective amount. A compoun 1 can be administered alone or as part of a pharmaceutically acceptable compositior . In addition, a compound or composition can be administered all at once, as for example by a bolus injection, multiple times, such as by a series of tablets, or delivered su bstantially uniformly over a period of time, as for example, using transdermal delivery. Fi irther, the dose of the compound can be varied over time. A compound can be administi ;red using an immediate release formulation, a controlled release formulation, or combinat ions thereof. The term "controlled release" includes sustained release, delayed rele ase, and combinations thereof.

[0039] A pharmaceutical composition of the invention can be prepared, packaged, or sold in bulk as a single unit dose, or as a plurality of single unit doses. As used herein, a "unit dose" s discrete amount of the pharmaceutical composition comprising a predetermin id amount of the active ingredient. The amount of the active ingredient is generally ec ual to the dosage of the active ingredient that would be administered to a patient or a convenient fraction of such a dosage such as, for example, one-half or one- third of sucl ι a dosage.

[0040] r ' he relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the human treated and further depe nding upon the route by which the composition is to be administered. By way of exar iple, the composition can comprise between 0.1 % and 100% (w/w) active ingredient. A unit dose of a pharmaceutical composition of the invention will generally comprise fr >m about 2 milligrams to about two grams of the active ingredient, and preferably omprises from about 10 milligrams to about 1 .0 gram of the active ingredient.

[0041] . Another aspect of the invention relates to a kit comprising a pharmaceutical compositiot of the invention and instructional material. Instructional material includes a publication, a recording, a diagram, or any other medium of expression which is used to communica :e the usefulness of the pharmaceutical composition of the invention for one of the purpc ses set forth herein in a human. The instructional material can also, for example, d< scribe an appropriate dose of the pharmaceutical composition of the invention. The instructional material of the kit of the invention can, for example, be

10

QB\1 1803388. 128833.00010 PCT Application affixed to a container which contains a pharmaceutical composition of the invention or be shipped tog rther with a container which contains the pharmaceutical composition.

Alternativel y, the instructional material can be shipped separately from the container with the intention that the instructional material and the pharmaceutical composition be used cooperative y by the recipient.

[0042] r Tie invention also includes a kit comprising a pharmaceutical composition of the inventio n and a delivery device for delivering the composition to a human. By way of example, the delivery device can be a squeezable spray bottle, a metered-dose spray bottle, an at rosol spray device, an atomizer, a dry powder delivery device, a self- propelling s alvent/powder-dispensing device, a syringe, a needle, a tampon, or a dosage- measuring c ontainer. The kit can further comprise an instructional material as described herein. The kit also comprises a container for the separate compositions, such as a divided boti le or a divided foil packet. Additional examples of containers include syringes, bc xes, bags, and the like. Typically, a kit comprises directions for the administrati on of the separate components. The kit form is particularly advantageous when the se arate components are preferably administered in different dosage forms (e.g., oral ai id parenteral), are administered at different dosage intervals, or when titration of the indiv dual components of the combination is desired by the prescribing physician.

[0043] ' . t may be desirable to provide a memory aid on the kit, e.g., in the form of numbers ne rt to the tablets or capsules whereby the numbers correspond with the days of the regimer that the tablets or capsules so specified should be ingested. Another example of such a m jmory aid is a calendar printed on the card, e.g., as follows "First Week, Monday, Ti lesday, . . . etc. . . . Second Week, Monday, Tuesday," etc. Other variations of memory aic s will be readily apparent. A "daily dose" can be a single tablet or capsule or several pill: or capsules to be taken on a given day.

[0044] n another embodiment of the present invention, a dispenser designed to dispense t : daily doses one at a time in the order of their intended use is provided. Preferably, the dispenser is equipped with a memory aid, so as to further facilitate compliance with the dosage regimen. An example of such a memory aid is a mechanical counter, wl ich indicates the number of daily doses that have been dispensed. Another example of such a memory aid is a battery-powered micro-chip memory coupled with a

1 1

QB\1 1803388. 128833.00010 PCT Application liquid crysti 1 readout, or audible reminder signal which, for example, reads out the date that the last daily dose has been taken and/or reminds one when the next dose is to be taken.

[0045] ' he compounds of the present invention, optionally comprising other pharmaceut cally active compounds, can be administered to a patient either orally, rectally, par ;nterally, (for example, intravenously, intramuscularly, or subcutaneously) intracistern; lly, intravaginally, intraperitoneally, intravesically, epidurally, otically, ophthalmic lly, locally (for example, powders, ointments or drops), or as a buccal or nasal spray. Other contemplated formulations include projected nanoparticles, liposomal preparation: , reseated erythrocytes containing the active ingredient, and

immunolog cally-based formulations.

[0046] 1 'arenteral administration of a pharmaceutical composition includes any route of administi ation characterized by physical breaching of a tissue of a human and administrati on of the pharmaceutical composition through the breach in the tissue.

Parenteral a dministration thus includes administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration includes subcutaneous, intraperiton jal, intravenous, intraarterial, intramuscular, or intrasterna! injection and intravenous , intraarterial, or kidney dialytic infusion techniques. For example, the compositions of the present invention can be administered to a subject by brain (via vPAG) inje ctions, intrathecal injections, intraperitoneal injections, or blood injections.

[0047] ' Compositions suitable for parenteral injection comprise the active ingredient combined vith a pharmaceutically acceptable carrier such as physiologically acceptable sterile aque )us or nonaqueous solutions, dispersions, suspensions, or emulsions, or may comprise st srile powders for reconstitution into sterile injectable solutions or dispersions. Examples c f suitable aqueous and nonaqueous carriers, diluents, solvents, or vehicles include wal sr, isotonic saline, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, ar d the like), suitable mixtures thereof, triglycerides, including vegetable oils such as oliv e oil, or injectable organic esters such as ethyl oleate. Proper fluidity can be maintained for example, by the use of a coating such as lecithin, by the maintenance of

12

QB\1 1803388. 128833.00010 PCT Application the required particle size in the case of dispersions, and/or by the use of surfactants. Such formulation i can be prepared, packaged, or sold in a form suitable for bolus

administrati an or for continuous administration. Injectable formulations can be prepared, packaged, o r sold in unit dosage form, such as in ampoules, in multi-dose containers containing < ·. preservative, or in single-use devices for auto-injection or injection by a medical pra ;titioner.

[0048] 1 ' ormulations for parenteral administration include suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradal le formulations. Such formulations can further comprise one or more additional ingredients including suspending, stabilizing, or dispersing agents. In one embodimen : of a formulation for parenteral administration, the active ingredient is provided in dry (i.e. powder or granular) form for reconstitution with a suitable vehicle (e.g. sterile ayrogen-free water) prior to parenteral administration of the reconstituted compositioi .

[0049] ' Ίιε pharmaceutical compositions can be prepared, packaged, or sold in the form of a st irile injectable aqueous or oily (emulsion) suspension or solution. This suspension >r solution can be formulated according to the known art, and can comprise, in addition ' o the active ingredient, additional ingredients such as the dispersing agents, wetting age its, or suspending agents described herein. Such sterile injectable formulatior 3 can be prepared using a non-toxic parenterally-acceptable diluent or solvent, such as wat ;r or 1 ,3-butanediol, for example. Other acceptable diluents and solvents include Rin jer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic m ano- or di-glycerides. Other parentally-administrable formulations which are useful inclu de those which comprise the active ingredient in microcrystalline form, in a liposomal p reparation, or as a component of a biodegradable polymer systems.

Compositio ns for sustained release or implantation can comprise pharmaceutically acceptable ] (olymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a spai ingly soluble polymer, or a sparingly soluble salt.

[0050] ' The compounds according to the present invention may also contain adjuvants such as pre; erving, wetting, emulsifying, and/or dispersing agents, including, for example, p; rabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be

13

QB\ 1 1803388. 128833.00010 PCT Application desirable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged a bsorption of injectable pharmaceutical compositions can be brought about by the use of agents capable of delaying absorption, for example, aluminum monostearate and/or gelat in. In particular, liposomes, mysomes and emulsifiers can be used in to make the pr jsent compounds more soluble for delivery.

[0051] Dosage forms can include solid or injectable implants or depots. In preferred embodimen ;s, the implant comprises an effective amount of an active agent and a biodegradal le polymer. In preferred embodiments, a suitable biodegradable polymer can be selected rom the group consisting of a polyaspartate, polyglutamate, poly(L-lactide), a poly(D,L- actide), a poly(lactide-co-glycolide), a poly(E-caprolactone), a

polyanhydride, a poly(beta-hydroxy-butyrate), a poly(ortho-ester) and a

polyphosph izene. In other embodiments, the implant comprises an effective amount of active agenl and a silastic polymer. The implant provides the release of an effective amount of a stive agent for an extended period of about one week to several years.

|0052] ! lolid dosage forms for oral administration include capsules, tablets, powders, and granule >. In such solid dosage forms, the active compound is admixed with at least one inert cu jtomary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, mannitol, or silicic acid; (b) bir ders, as for example, carboxymethylcellulose, alginates, gelatin, polyvinylp) rrolidone, sucrose, or acacia; (c) humectants, as for example, glycerol; (d) disintegratii ig agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, algii ic acid, certain complex silicates, or sodium carbonate; (e) solution retarders, as for exam Die, paraffin; (f) absorption accelerators, as for example, quaternary ammonium compounds; (g) wetting agents, as for example, cetyl alcohol or glycerol monosteara e; (h) adsorbents, as for example, kaolin or bentonite; and/or (i) lubricants, as for exampli , talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium laui yl sulfate, or mixtures thereof. In the case of capsules and tablets, the dosage forms may ilso comprise buffering agents.

[0053] . ^ tablet comprising the active ingredient can, for example, be made by compressin i or molding the active ingredient, optionally with one or more additional ingredients, Compressed tablets can be prepared by compressing, in a suitable device,

14

QB\I 1803388. 128833.000 10 PCT Application the active ir gredient in a free-flowing form such as a powder or granular preparation, optionally mixed with one or more of a binder, a lubricant, an excipient, a surface active agent, and a dispersing agent. Molded tablets can be made by molding, in a suitable device, a mi xture of the active ingredient, a pharmaceutically acceptable carrier, and at least sufficient liquid to moisten the mixture.

[0054] I 'harmaceutically acceptable excipients used in the manufacture of tablets include iner : diluents, granulating and disintegrating agents, binding agents, and lubricating ι .gents. Known dispersing agents include potato starch and sodium starch glycolate. Known surface active agents include sodium lauryl sulfate. Known diluents include calc ium carbonate, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, < alcium hydrogen phosphate, and sodium phosphate. Known granulating and disintegratii g agents include corn starch and alginic acid. Known binding agents include gelatin, aca< ia, pre-gelatinized maize starch, polyvinylpyrrolidone, and hydroxypropyl methylcellu ose. Known lubricating agents include magnesium stearate, stearic acid, silica, and talc.

[0055] r ' ablets can be non-coated or they can be coated using known methods to achieve deli yed disintegration in the gastrointestinal tract of a human, thereby providing sustained re lease and absorption of the active ingredient. By way of example, a material such as glyc eryl monostearate or glyceryl distearate can be used to coat tablets. Further by way of e cample, tablets can be coated using methods described in U.S. Pat. Nos. 4,256,108; ' -,160,452; and 4,265,874 to form osmotically-controlled release tablets. Tablets can further comprise a sweetening agent, a flavoring agent, a coloring agent, a preservative , or some combination of these in order to provide pharmaceutically elegant and palatab e preparation.

[0056] ! >olid dosage forms such as tablets, dragees, capsules, and granules can be prepared wi th coatings or shells, such as enteric coatings and others well known in the art. They n ay also contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a delayed manner. Examples of embedding ;ompositions that can be used are polymeric substances and waxes. The active comp ounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

15

QB\ 1 1803388.1 128833.00010 PCT Application

[0057] S lolid compositions of a similar type may also be used as fillers in soft or hard filled gelatin capsules using such excipients as lactose or milk sugar, as well as high molecular v eight polyethylene glycols, and the like. Hard capsules comprising the active ingredient c in be made using a physiologically degradable composition, such as gelatin. Such hard c ipsules comprise the active ingredient, and can further comprise additional ingredients ncluding, for example, an inert solid diluent such as calcium carbonate, calcium phc sphate, or kaolin. Soft gelatin capsules comprising the active ingredient can be made usi ng a physiologically degradable composition, such as gelatin. Such soft capsules comprise the active ingredient, which can be mixed with water or an oil medium such as peanut oil, liquid paraffin, or olive oil.

[0058] ( )ral compositions can be made, using known technology, which specifically release oral y-administered agents in the small or large intestines of a human patient. For example, fo -mulations for delivery to the gastrointestinal system, including the colon, include enU ric coated systems, based, e.g., on methacrylate copolymers such as poly(methai irylic acid, methyl methacrylate), which are only soluble at pH 6 and above, so that the p olymer only begins to dissolve on entry into the small intestine. The site where such polymer formulations disintegrate is dependent on the rate of intestinal transit and the amc unt of polymer present. For example, a relatively thick polymer coating is used for del ivery to the proximal colon (Hardy et al., Aliment. Pharmacol. Therap. (1987) 1 :273-280). Polymers capable of providing site-specific colonic delivery can also be used, where in the polymer relies on the bacterial flora of the large bowel to provide enzymatic c egradation of the polymer coat and hence release of the drug. For example, azopolymei s (U.S. Pat. No. 4,663,308), glycosides (Friend et al., J. Med. Chem. (1984) 27:261 -268 1 and a variety of naturally available and modified polysaccharides (see PCT application PCT/GB89/00581 ) can be used in such formulations.

[0059] 'ulsed release technology such as that described in U.S. Pat. No. 4,777,049 can also be used to administer the active agent to a specific location within the gastrointest nal tract. Such systems permit drug delivery at a predetermined time and can be used to c eliver the active agent, optionally together with other additives that my alter the local m: croenvironment to promote agent stability and uptake, directly to the colon,

16

QB\1 1803388. 128833.00010 PCT Application without rely ing on external conditions other than the presence of water to provide in vivo release.

[0060] I liquid dosage forms for oral administration include pharmaceutically acceptable e mulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage form may contain inert diluents commonly used in the art, such as wati ;r or other solvents, isotonic saline, solubilizing agents and emulsifiers, as for example, et yl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benz Date, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils, in particular, a lmond oil, arachis oil, coconut oil, cottonseed oil, groundnut oil, corn germ oil, olive oil , castor oil, sesame seed oil, MIGLYOL™, glycerol, fractionated vegetable oils, minera oils such as liquid paraffin, tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid es ters of sorbitan, or mixtures of these substances, and the like.

[0061] I tesides such inert diluents, the compounds of the present invention can also include adji vants, such as wetting agents, emulsifying and suspending agents, demulcents, preservatives, buffers, salts, sweetening, flavoring, coloring and perfuming agents. Sus sensions, in addition to the active compound, may contain suspending agents, as for exam >le, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol or sorbitan esters, micr >crystalline cellulose, hydrogenated edible fats, sodium alginate, polyvinylp> rroIidone, gum tragacanth, gum acacia, agar-agar, and cellulose derivatives such as sod urn carboxymethylcel!ulose, methylcellulose, hydroxypropylmethylcellulose, aluminum r letahydroxide, bentonite, or mixtures of these substances, and the like.

Liquid fonr ulations of a pharmaceutical composition of the invention that are suitable for oral admini :tration can be prepared, packaged, and sold either in liquid form or in the form of a di y product intended for reconstitution with water or another suitable vehicle prior to use

[0062] ] nown dispersing or wetting agents include naturally-occurring phosphatides such as leci hin, condensation products of an alkylene oxide with a fatty acid, with a long chain aliphi tic alcohol, with a partial ester derived from a fatty acid and a hexitol, or with a partial est ;r derived from a fatty acid and a hexitol anhydride (e.g. polyoxyethylene stearate, he] rtadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyeth; 'lene sorbitan monooleate, respectively). Known emulsifying agents include

17

QB\1 1803388. 128833.000 10 PCT Application lecithin and acacia. Known preservatives include methyl, ethyl, or n-propyl-para- hydroxyben roates, ascorbic acid, and sorbic acid. Known sweetening agents include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin. Known thickening agents for o ly suspensions include, for example, beeswax, hard paraffin, and cetyl alcohol.

[0063] I ,iquid solutions of the active ingredient in aqueous or oily solvents can be prepared in substantially the same manner as liquid suspensions, the primary difference being that tl e active ingredient is dissolved, rather than suspended in the solvent. Liquid solutions of the pharmaceutical composition of the invention can comprise each of the components described with regard to liquid suspensions, it being understood that suspending igents will not necessarily aid dissolution of the active ingredient in the solvent. Aq ueous solvents include, for example, water and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesan e, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.

[0064] ( compositions of the present invention may further include a cyclodextrin component n order to, e.g., improve water solubility of an active pharmaceutical ingredient, ] irolong drug release, and improve tabletting characteristics. In general, cyclic structure oli gomers of glucose ("cyclodextrin") are obtained from the starch digests of certain bact :ria. The most abundant cyclodextrins are alpha, beta and gamma cyclodextri which have 6,7 and 8 glucose units, respectively. The interior cavity of a cyclodextrin is hydrophobic and the exposed surface of the molecule is hydrophilic. Cyclodextri is are known to enhance active pharmaceutical ingredient stability, aqueous solubility, a id reduce volatility. Some examples of commercially available cyclodextrin, or derivativ ;s thereof, are as follows: alpha-Cyclodextrin (CAS #: 10016-20-3); (2- Hydroxyprc pyl)-alpha-cycIodextrin (CAS #: 128446-33-3); beta-Cyclodextrin (CAS #: 7585-39-9); 6-O-alpha-D-Glucosyl-beta-cyclodextrin (CAS #: 92517-02-7); gamma- Cyclodextrn (CAS #: 17465-86-0); and (2-Hydroxypropyl)-gamma-cyclodextrin (CAS #: 128446-24-4). Cyclodextrins particularly useful in formulating a delivery vehicle to administer 1 he present compounds include: the sulfobutyl ether beta-cyclodextrin (SBE- beta-CD) a\ ailable from Cydex Pharmaceuticals, Inc. under the tradename CAPT1SOL;

18

QB\1 1803388.1 128833.000 10 PCT Application and the cyc dextrin and hydroxypropyl betacyclodextrins available from Roquette Pharma und ΪΓ the tradename LEPTOSE. An exemplary intravenous formulation may be formulati :d in saline water containing 0.9% sodium chloride, 450 mg/mL of

CAPTISOL cyclodextrin, and 1 .5 mg/mL of the compound designated CMX-020 described ai d claimed herein.

[0065] ( 'ompositions for rectal or vaginal administration can be prepared by mixing a compound c f the present invention and any additional compounds with suitable non- irritating ex :ipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ordinary room temperature, but liquid at body temperature, and therefore, rr elt in the rectum or vaginal cavity and release the active ingredient. Such a compositior can be in the form of, for example, a suppository, a retention enema preparation, and a solution for rectal or colonic irrigation. Suppository formulations can further com )rise various additional ingredients including antioxidants and preservatives. Retention ei lema preparations or solutions for rectal or colonic irrigation can be made by combining 1 lie active ingredient with a pharmaceutically acceptable liquid carrier. As is known in th ; art, enema preparations can be administered using, and can be packaged within, a de ivery device adapted to the rectal anatomy of a human. Enema preparations can further i :omprise various additional ingredients including antioxidants and preservative s.

[0066] pharmaceutical composition of the invention can be prepared, packaged, or sold in a fot mulation suitable for vaginal administration. Such a composition can be in the form of, for example, a suppository, an impregnated or coated vaginally-insertable material sue h as a tampon, a douche preparation, or a solution for vaginal irrigation.

[0067] 1 )osage forms for topical administration of a compound according to the present invt ntion include ointments, powders, sprays and inhalants. The compounds are admixed un ier sterile conditions with a physiologically acceptable carrier, and any preservative s, buffers, and/or propellants that may be required. Formulations suitable for topical adm nistration include liquid or semi-liquid preparations such as liniments, lotions, oil- n-water or water-in-oil emulsions such as creams, ointments or pastes, and solutions or suspensions. Topically-administrable formulations can, for example, comprise fr >m about 0.1 % to about 10% (w/w) active ingredient, although the

19

QBM 1 803388. 1 128833.00010 PCT Application concentratic n of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration can further comprise one or more of the additional ingredients described herein.

[0068] ( )phthalmic formulations, eye ointments, powders, and solutions are also contemplate d as being within the scope of this invention. Such formulations can, for example, be in the form of eye drops including, for example, a 0.1 -1 .0% (w/w) solution or suspensic n of the active ingredient in an aqueous or oily liquid carrier. Such drops can further com >rise buffering agents, salts, or one or more other of the additional ingredients described herein. In other embodiments, ophtha!malmically administrable formulations comprise th : active ingredient in microcrystalline form or in a liposomal preparation.

[0069] I 'harmaceutical compositions of the invention formulated for pulmonary delivery car provide the active ingredient in the form of droplets of a solution or suspension. Such formulations can be prepared, packaged, or sold as aqueous or dilute alcoholic so lutions or suspensions, optionally sterile, comprising the active ingredient, and can con veniently be administered using any nebulization or atomization device. Such formu ations can further comprise one or more additional ingredients including a flavoring ag ent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agenl , or a preservative such as methyl hydroxybenzoate. The droplets provided by this routi : of administration preferably have an average diameter in the range from about 0.1 to about 200 nanometers.

[0070] pharmaceutical composition of the invention can be prepared, packaged, or sold in a foi mulation suitable for buccal administration. Such formulations can, for example, be in the form of tablets or lozenges made using conventional methods, and can, for exa nple, comprise 0.1 to 20% (w/w) active ingredient, the balance comprising an orally di: solvable or degradable composition and, optionally, one or more of the additional ii igredients described herein. Alternately, formulations suitable for buccal administrati on can comprise a powder or an aerosolized or atomized solution or suspension ;omprising the active ingredient. Such powdered, aerosolized, or atomized formulation 5, when dispersed, preferably have an average particle or droplet size in the range from ibout 0.1 to about 200 nanometers, and can further comprise one or more of the addition al ingredients described herein.

20

QB\ 1 1803388. 1 128833.00( Ί 0 PCT Application

[0071] " or parenteral administration in non-human animals, the compounds of the present inv :ntion may be prepared in the form of a paste or a pellet and administered as an implant, usually under the skin of the head or ear of the animal. Paste formulations can be preparec by dispersing a compound or compounds in pharmaceutically acceptable oil such as pea nut oil, sesame oil, corn oil or the like. Pellets containing a therapeutically effective ar iount of a compound or compounds can be prepared by admixing the compound A'ith a diluent such as a carbowax, carnauba wax, and the like, and a lubricant, such as ma; mesium or calcium stearate, can be added to improve the pelleting process. It is, of coursi recognized that more than one pellet may be administered to an animal to achieve the desired dose level. Moreover, it has been found that such implants may also be adminisi ered periodically during the animal treatment period in order to maintain the proper acti v e agent level in the animal's body.

[0072] The compounds of the present invention and the pharmaceutically acceptable salts of the same, can be administered to a patient at dosage levels in the range of from about 0.01 o about 1 ,000 mg per day. For a normal adult human having a body weight of about 70 kg, a dosage in the range of from about 0.01 to about 300 mg is typically sufficient, with 1 -10 mg a preferred dosage. However, some variability in the general dosage ranj ;e may be required depending upon the age and weight of the subject being treated, the intended route of administration, the particular compound being administered and the like . The determination of dosage ranges and optimal dosages for a particular patient is w ell within the ability of one of ordinary skill in the art having the benefit of the instant disc losure. It is also noted that the compounds of the present invention can be used in sus ained release, controlled release, and delayed release formulations, which forms are a !so well known to one of ordinary skill in the art.

[0073] It is not critical whether the compounds of the present invention are administers d directly to the cell, to a tissue comprising the cell, a body fluid that contacts the cell, or i body location from which the compound can diffuse or be transported to the cell. It is si ifficient that the compound is administered to the patient in an amount and by a route wh« reby an amount of the compound sufficient to mobilize lipids in the cell arrives, diructly or indirectly at the cell. The minimum amount varies with the identity of the compoi nds.

21

QB\1 1803388. 128833.00010 PCT Application

[0074] The specific dosage and dosage range that can be used depends on a number of factors, i ncluding the requirements of the patient, the severity of the condition being treated, anc the pharmacological activity of the compound being administered. The determinati on of dosage ranges and optimal dosages for a particular patient is well within the ordinar > skill of one in the art in view of this disclosure. It is understood that the ordinarily s killed physician, dentist, or veterinarian will readily determine and prescribe an effectivi amount of the compound to mobilize lipid stores, induce weight loss, or inhibit apputite in the patient. In so proceeding, the physician or veterinarian can, for example, p escribe a relatively low dose at first, subsequently increasing the dose until an appropriate response is obtained. It is further understood, however, that the specific dose level for an y particular human will depend upon a variety of factors including the activity of the speci fic compound employed, the age, body weight, general health, gender, and diet of the human, the time of administration, the route of administration, the rate of excretion, ι ny drug combination, and the severity of any disorder being treated.

[0075] The compounds of the present invention are particularly useful when formulated in the form of a pharmaceutical injectable dosage, including a compound described a nd claimed herein in combination with an injectable carrier system. As used herein, inje ctable and infusion dosage forms (i.e., parenteral dosage forms) include, but are not lim ited to, liposomal injectables or a lipid bilayer vesicle having phospholipids that encapsulate an active drug substance. Injection includes a sterile preparation intended fo parenteral use.

[0076] ^ive distinct classes of injections exist as defined by the USP: emulsions, lipids, pow ders, solutions and suspensions. Emulsion injection includes an emulsion comprising a sterile, pyrogen-free preparation intended to be administered parenterally. Lipid com] »lex and powder for solution injection are sterile preparations intended for reconstituti jn to form a solution for parenteral use. Powder for suspension injection is a sterile prep aration intended for reconstitution to form a suspension for parenteral use. Powder lyc philized for liposomal suspension injection is a sterile freeze dried preparation intended fc r reconstitution for parenteral use that is formulated in a manner allowing incorporation of liposomes, such as a lipid bilayer vesicle having phospholipids used to encapsulate an active drug substance within a lipid bilayer or in an aqueous space,

22

QB\1 1803388. 128833.00010 PCT Application whereby ti e formulation may be formed upon reconstitution. Powder lyophilized for solution in ection is a dosage form intended for the solution prepared by lyophilization ("freeze dr ying"), whereby the process involves removing water from products in a frozen stat i at extremely low pressures, and whereby subsequent addition of liquid creates a sc lution that conforms in all respects to the requirements for injections. Powder lyophilized for suspension injection is a liquid preparation intended for parenteral use that contains sc lids suspended in a suitable fluid medium, and it conforms in all respects to the require nents for Sterile Suspensions, whereby the medicinal agents intended for the suspension are prepared by lyophilization. Solution injection involves a liquid preparation containing one or more drug substances dissolved in a suitable solvent or mixture of mutually miscible solvents that is suitable for injection. Solution concentrate injection ir volves a sterile preparation for parenteral use that, upon addition of suitable solvents, y elds a solution conforming in all respects to the requirements for injections. Suspension injection involves a liquid preparation (suitable for injection) containing solid particles di spersed throughout a liquid phase, whereby the particles are insoluble, and whereby s n oil phase is dispersed throughout an aqueous phase or vice-versa. Suspension liposomal injection is a liquid preparation (suitable for injection) having an oil phase d spersed throughout an aqueous phase in such a manner that liposomes (a lipid bilayer ves icle usually containing phospholipids used to encapsulate an active drug substance < ither within a lipid bilayer or in an aqueous space) are formed. Suspension sonicated njection is a liquid preparation (suitable for injection) containing solid particles di spersed throughout a liquid phase, whereby the particles are insoluble. In addition, the product may be sonicated as a gas is bubbled through the suspension resulting in the formation of microspheres by the solid particles.

[0077] The parenteral carrier system includes one or more pharmaceutically suitable excipients, such as solvents and co-solvents, solubilizing agents, wetting agents, suspending agents, thickening agents, emulsifying agents, chelating agents, buffers, pH adjusters, a itioxidants, reducing agents, antimicrobial preservatives, bulking agents, protectants tonicity adjusters, and special additives.

[0078] The invention further contemplates formulating combination pharmaceutical compositio is which include both a compound as described and claimed herein and an

23

QB\ 1 1803388. 128833.00C 10 PCT Application anesthetic £ gent. Such compositions are useful in medical procedures including, but not limited to, ] ;eneral anesthesia, sedation for mechanically ventilated subjects, and procedural sedation. In general, an "anesthetic agent" is a drug that brings about a state of anesthes a in a subject. However, while many current anesthetic agents produce unconsciou mess, they provide no analgesia and must be used in combination with other drugs. For sxample, propofol is approved in more than fifty countries, and generic versions arc available. Propofol is regularly administered in combination with opioids, such as fen anyl, alfentanil, remifentanii and sufentanil, to provide combination hypnotic effect and f ain alleviation. As can be appreciated, compounds of the present invention are suitable for replacing such opioid analgesics in combination anesthetic/analgesic formulatior s and for use in related medical procedures. A variety of anesthetic agents may be use 1 in combination with the present compounds, including intravenous agents such as barbiturates, benzodiazepines, etomidate, ketarnine and propofol. In one embodimer t, the analgesic compound designated CMX-020 is combined in a pharmaceut ical composition with propofol, (available from Astra Zeneca under the tradename ! DIPRIVAN) to provide an intravenously administered formulation providing both hypno ;ic and analgesic effects.

[0079] Various exemplary embodiments of compositions and methods according to this inventi >n are now described in the following examples. In these embodiments, specific pro ducts identified by Arabic numerals (e.g., 1 , 2, 3, etc.) refer to the specific structures S 3 identified in the following description. The following examples are offered for illustrat ve purposes only and are not intended to limit the scope of the present invention ii i any way. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing d escription and the following examples and fall within the scope of the appended c aims.

III . EXAMPLES

Example 1 : Characterization of CMX-020.

[0080] The present example describes the characterization of compound CMX-020, which is de Dieted in Fig. 2, as is AM-404.

24

QB\1 1 803388. 128833.00( 10 PCT Application

[0081] CMX-020 Binding Assay Results. CMX-020 was tested in the Cerep Full BioPrint Pi ofile, which is a panel of 158 different in vitro receptor binding and enzyme assays. Fc r the initial BioPrint screen, a free compound concentration of 10 μΜ was used, whicl i corresponds to a 2.5mg/kg bolus dose in a mouse. When injected the CMX- 020 lipid therapeutic becomes partially bound to plasma or serum proteins; our analysis also shows approximately 99% of CMX-020 is bound. Thus, a 2.5 mg/kg bolus dose would be e mivalent to a 0.1 μΜ free compound concentration. For the BioPrint Assay, any receptc r binding or enzyme assays that display inhibition over 50%, at a CMX-020 concentrati )n of 10 μΜ, was singled out for further IC50 analysis. Table 1 below shows all receptor s in the Cerep BioPrint Profile with inhibition over 50% along with their IC50 concentrati >ns.

Table 1.

CM) [-020 - Assay Binding Summary (BioPrint/Cerep)

[0082] Receptors Implicated in Dependence and Addiction. Of the 158 receptors and enzym ;s screened in the BioPrint assay, 53 are selected by Cerep for implication in the depend :nce and addiction panel (see receptor and enzyme category table below). Out of the 53 ri jceptors and enzymes implicated in dependence and addiction, CMX-020 had significant inhibition in only the cannabinoid and opioid receptors. The role of these receptors ir dependence and addiction is discussed below.

[0083] CB1 Receptor in Dependence and Addiction. It is well known that natural ligands for the CB 1 cannabinoid receptors, anandamide and 2-arachidonylglycerol, affect nervous sy item functions such as reward, memory, cognition, and pain perception. The

25

QB\1 1803388. 1 128833.001 ) 10 PCT Application perceived : ole of CB 1 receptors in dependence and addiction is driven largely by Δ9- THC, the ictive ingredient in marijuana, which is a popular recreational drug. It is believed tl at A9-THC produces dependency or addiction in two ways: by mimicking natural lig ands for CBl receptors and by producing elevated dopamine levels. In comparing dependence and addiction with other drugs however, marijuana does not have the same 1 ;vel of risk as opioids (e.g., morphine, heroin), cocaine, or alcohol. Unlike opioids, cc caine and alcohol, there is little risk in overdosing on marijuana. Also, the developme it of dependence on marijuana is much less prevalent than nicotine and cocaine. W hile CMX-020 is a strong CB l receptor agonist, it is unlike A9-THC in that it does not el ;vate dopamine levels. CMX-020 is more like the active ingredient in Tylenol, AM404, w n ' ch is both a CB l and TRPV 1 agonist, and which has no significant risk for dependence and addiction.

[0084] K-Opioid and δ-Opioid in Dependence and Addiction. Opioid receptors, μ, K, δ (mu, k ippa, and delta), are G protein-coupled receptors found in the central nervous system. M ost traditional opioids used in pain management, like morphine and fentanyl, as well a s the highly addictive opioid heroin, are μ-opioid receptor agonists. Interestingl y, morphine's addictive properties are completely abolished in mice lacking the μ-opioi i receptor. Thus, the μ-opioid receptor is responsible for the addictive effects of morphir e and most other traditional opioids used in pain management. As the CMX- 020 assay 1 finding table above shows, CMX-020 is an active agonist of the -opioid and δ-opioid re ceptors, but not the μ-opioid receptor. The role of the δ-opioid receptor in reward and addiction is still poorly understood, but there is emerging evidence that the δ- opioid rece ptor is involved in opioid reward and addiction. Accordingly, CMX-020 is a relatively veak δ-opioid agonist - its IC50 concentration as a δ-agonist is nearly 3x lower than its va ue as a δ-agonist. But moreover, the κ-opioid receptor activation has been shown to p roduce aversive states, which should suppress any low-level, but unwanted δ- mediated ri sks of dependence and addiction.

[0085] The screening results of CMX-020 in the full BioPrint Profile (Cerep) provides ir lportant information on dependence and addiction. The primary concern for CMX-020 nay be the cannabinoid and opioid agonist activity. Although CMX-020 is a strong CB l agonist, it does not elevate dopamine levels like A9-THC. Its combination of

26

QB\I 1803388. 1 CB1 and TR V1 activity makes it similar to AM404, the active ingredient in Tylenol which is proven to show no apparent risk for dependence and addiction. CMX-020 also shows agonist activity for the κ-opioid and ό -opioid receptors. This combination appears to constitute a unique non-habit forming alternative to traditional μ-opioids for the treatment of pain.

|O086] Table 2 below presents the Cerep BioPrint receptors and enzymes that are implicated in dependence and addiction, have been screened using CMX-020, but are not active.

Table 2

Receptors and Enzymes Implicated in Addiction

but Not Active for CMX-020

2? 128833.001110 PCT Application

Example 2 : Analgesic Effect of CMX-020 and Intravenous/Oral Formations.

[0087] Based on the inventors' research, CMX-020, is a rapidly acting intravenous analgesic that can be used as an adjunct to traditional opioids or as a stand-alone analgesic. Chemically, it is close structural analog of a set of lipid mediators derived from arach donic acid that the body uses to control pain. Like acetaminophen, the analgesic e Tect of CMX-020 also appears to be produced through the CBI and TRPVl receptors. Side-by-side comparisons of AM404 with CMX-020 demonstrate that CMX- 020 is a mc re potent analgesic without the toxic effects. Because CMX-020 does not have the to cic effects of acetaminophen, higher daily doses are possible. When higher doses are u >ed, the analgesic effect is similar to morphine.

[0088] MX-020 is very slightly soluble in water and, thus, is formulated in an opaque wh te, oil-in-water emulsion. When produced in bulk quantities, it contains 1 Omg/ml o * CMX-020, described here. In addition to the active component, CMX-020, a preferred is otonic formulation also contains, by weight, soybean oil (10%), glycerol (2.25%), T veen 80 (0.61%), hydrogenated phospholipid (0.49%) and disodium edetate (0.005%); with sodium hydroxide to adjust pH. CMX-020 provided as an Injectable Emulsion i: isotonic and has a pH of 7-8.5. The structural formula for CMX-020 is, C26H44N2O .:

Molecular ' veight: 416.64

[0089] The precise mechanism of the analgesic properties of CMX-020 has yet to be established and certainly no one mode of operation is adopted herein. However, like acetaminop hen, the effects of CMX-020 appear to be mediated, in part, through cannabinoil i and vanilloid receptors. Studies with CBi cannabinoid and TRPVl vanilloid knockout n ice (both homozygotes) show that the effect of CMX-020 is altered in both; but in CBi ;annabinoid knockout mice, the analgesic effect of CMX-020 is almost completely blocked. Furthermore, the mechanisms appear unique and different from morphine, l entanyl, and NSAIDs.

28

QB\1 1803388. 128833.00 ) 10 PCT Application

[0090] CMX-020 provided as the Injectable Emulsion (l mg/mL) is a fast-acting intravenou ι pain therapeutic that performs like morphine, providing high levels of analgesia, 1 vhich can be established through dose titration. CMX-020 is apparently a more poter t analgesic than acetaminophen, without the API chemistry that makes acetaminof hen toxic to the liver. Accordingly, CMX-020 can function as an alternative to acetaminof hen, an opioid-sparing therapeutic, or as an opioid replacement. Intravenous injection o: ' a therapeutic dose of CMX-020 produces analgesia rapidly, usually within 3- 5 minutes i rom the start of an injection. As with other rapidly acting intravenous agents, the half-tin ie of the blood-brain equilibration is approximately 1 to 3 minutes, and this accounts fc r the rapid induction of analgesia.

[0091] CMX-020 provided as the Injectable Solution provides onset of pain relief within 3-5 ninutes after bolus injection or infusion and is appropriate for mild, moderate, or severe a ;ute pain. The duration of effect from bolus injection is 20-30 minutes. A sustained a ialgesic effect is established using continuous infusion. The level of analgesic effect can I e adjusted through dose titration. Sustained levels of analgesia can be maintained for 48 hours or more. Specific applications include: acute post operative pain manageme it; breakthrough pain therapy; intensive care units; acute trauma; intravenous patient-con trolled analgesia; and end of life pain control.

[0092] CMX-020 Injectable Solution may also be given to patients during surgery or dental proc sdures to relieve pain and as an adjunct to an anaesthetic. CMX-020 Injectable Solution al io reduces fever within 30 minutes after the start of administration with duration of the antipyretic effect of at least 2 hours after ending administration.

[0093] Dosage and Analgesic Equivalence with Acetaminophen and Morphine has been deterr lined by the inventors for CMX-020. Using the preclinical writhing assay in mice to establish relative levels of analgesia, the dose of CMX-020 that provides an equivalent l x) level of analgesia to the recommended dose of intravenous

acetaminof: hen (the equivalent of 1 ,000 mg in human) is a bolus injection of 0.01 mg/kg, followed by continuous infusion at a rate of 0.08 mg/kg/hr. The level of analgesia established by acetaminophen is not dose dependent. The level of analgesia established by CMX-0: 10 is dose dependent. The doses of CMX-020 that correspond to higher levels

29

QB\1 1803388. 1 128833.00010 PCT Application of analgesi ι in comparison to acetaminophen in the mouse writhing assay (2x and 3x) are shown in T able 3 below.

[0094] Table 3. Dose Equivalence and Maximum Recommended Doses of CMX- 020 Injecta ole Emulsion

Maximum Recommended LD50**

CMX-020 Doses CMX-020 Doses

0.08 mg/kg 2.4 mg/kg

0.64 mg kg/hr 6.4 mg kg/hr

For a 70 kg human: For a 70 kg human:

5.6 mg 170 mg 45 mg/hr 450 mg/hr

Table 3.

[0095] LD50 (abbreviation for "Lethal Dose, 50%") is established using mice. The dose translation between mouse and human is based on body surface area allometric translation. For example, a 1 mg/kg mouse dose is equivalent to a 0.08 mg/kg human dose. For ι nore information on species dose translation see: Reagan-Shaw et.al, FASEB J.22, 659-651 (2007).

Table 4. Analgesic Equivalence

for a 70 kg human: Dosing for

CMX-020 and Morphine

[0096] CMX-020 can also be given in doses that provide equivalent analgesic levels to different doses of morphine, as shown in Table 4 above. For bolus injections, the analgesic e Tect of CMX-020 is equivalent to morphine on a mg/kg basis. For example, a 1 mg bolus dose of CMX-020 will establish roughly the same level of analgesia as a 1 mg bolus dose af morphine. However, CMX-020 will be faster acting (within 3-5 minutes for CMX-C 20 versus 20 minutes for morphine) and its analgesic effect will be shorter in duration (2 )-25 minutes for CMX-020 versus 35-40 minutes for morphine). To maintain a constant l evel of analgesia, a bolus injection should be followed by continuous infusion.

30

QB\1 1803388. 1 128833.001 ) 10 PCT Application

To establis ι an analgesic equivalence with morphine via continuous infusion, twice the mg/kg dos< ! of CMX-020 is required. For example, a 6mg/hr infusion dose of CMX-020 is equivale it to a 3mg/hr infusion dose of morphine. Like morphine, the dose of CMX- 020 can be titrated over a wide range (see Table 3 above) to establish the patient required analgesic level.

Example 3 : AA analog efficacy in standard pain assays.

[0097] In this example, the inventors compare the performance of intravenously delivered C MX-020 to both Perfalgan (intravenous acetaminophen) and morphine (the market leac ing intravenous opioid) in both the tail-flick and writhing assay. The tail-flick assay repre sents the most severe acute pain indications. The writhing assay is a more moderate p lin assay that represents internal noxious pain indications, but also encompass ;s inflammatory, chemical, and persistent central pain indications. An important c ifference between the two assays is the strength of analgesic required to be effective in each of the assays. As seen below, to achieve a moderate level of efficacy, the tail-flic c assay requires roughly l OOx higher dose than the writhing assay from both CMX-020 i ind morphine. The administration of assays is described below.

[0098] Tail-Flick Assay in Mouse: The tail-flick assay is based on the time measured to reflex wi thdrawal of the tail in response to a radiant heat source. The maximum exposure ti ne to the radiant heat source is set at 10 seconds. Prior to treatment, a baseline (B L) time measured to tail-flick withdrawal is determined after two exposures to the radiant source separated by a 30 min period. Control mice are intravenously treated with vehicl ; and test mice are intravenously treated with analgesic test compound. Data of tail-flick withdrawal of treated mice (TM) are calculated as maximum possible effect (MPE), wh ;re MPE = (TM-BL)/(10-BL). Tail-flick withdrawal is measured at 5 min, 15 min, 30 mil i, and 1 hr. A total of 3-5 mice are used for each time point tested.

[0099] Writhing Assay in Mouse: The writhing assay uses an injection of dilute acetic acid Ό.55%) intraperitoneal ly which stimulates an internal pain response that results in w rithing of the mouse. A writhe is indicated as a whole body stretch or contraction of the abdomen. The mean number of writhes is counted over 5 min periods between 5- 0 min, 15-20 min, 25-30 min, and 35-40 minutes after intravenous injection

31

QB\1 1803388. 128833.00( Ί 0 PCT Application of either a 1 est analgesic for treated mice (TM) or vehicle for baseline (BL). At each time point, data if writhing assay are calculated as maximum possible effect (MPE), where MPE = (1 - ' ^ M/BL)* 100. A total of 5 mice are used for each analgesic tested.

[00100] Tail-flick Assay Performance Comparison - Bolus Injection. In Figure 4, the dose re! ponse of CMX-020 is delivered in its emulsion vehicle and compared with morphine; ; md the two compounds show very similar analgesic potency at the same doses. The time course comparison in Figure 4 shows that CMX-020 acts very quickly, providing a maximal analgesic response at 5 minutes. The duration of this maximal CMX-020 i esponse lasts through 30 minutes with a 10 mg/kg dose. At the same dose, morphine ti ikes 15 minutes to establish its full analgesic effect. This effect also lasts through 30 minutes.

[00101] Writhing Assay Performance Comparison - Bolus Injection. In the first graph of Fi jure 5, the dose response of CMX-020 and morphine again show similar potency an< I both compounds achieve 100% MPE at a dose of approximately 0.5 mg/kg. With Perfal an (intravenous acetaminophen), a 12.5 mg/kg dose establishes a relatively low, but m< asurable analgesic response of approximately 35% MPE. While only a single dose of Per Falgan is shown in the first graph, our tests show that the level of analgesia provide by Perfalgan does not increase above 35% MPE in the writhing assay for doses up to 200m g/kg. At a dose of 0.05 mg/kg, CMX-020 provides a higher analgesic response th in Perfalgan at a 12.5 mg/kg dose; here, the dose of Perfalgan is 250 times that of CMX-020.

[00102] Infusion Studies. For patients requiring longer-term pain management, the continuous infusion of pain therapeutics enables the maintenance of constant levels of analgesia for long periods. In continuous infusion, a therapeutic is delivered in an infusion fli id at a constant flow rate through a peripheral venous catheter. The flow rate of infusion fluids can be controlled by either adjusting the drip rate, or more precisely, by using an in usion pump that also controls the total therapeutic dose delivered. In the second graph of Figure 5, mice received an infusion of either CMX-020 or morphine over a 24 hour p jriod. The infusion of CMX-020 and morphine utilizes a miniature therapeutic Alzet pump that is connected to a surgically implanted jugular vein cannula. This Alzet sump, which is implanted under the skin, administers a constant flow of

32

QB\ 1 1803388. 128833.00M0 PCT Application therapeutic over 24 hours. Using the writhing assay, analgesic response levels are measured a . 3 hours and 20 hours after beginning the continuous infusion. Doses of both CMX-020 ; ind morphine were chosen so that analgesia levels were produced at roughly 50% MPE n the writhing assay. Figure 5 shows preliminary results that utilize three to four anima! s for each time point and for controls. Several important conclusions can be made from this study. First, analgesia can be sustained at constant levels over a 24 hour period usin continuous infusion. Analgesia levels in the second graph of Figure 5 are greater that those provided by a 12.5 mg/kg dose of Perfalgan (acetaminophen). Second, the dose of CMX-020 required to maintain the approximate level of analgesia as morphine i; only slightly higher: 2 mg/kg/hr of CMX-020 produces roughly the same level of an. lgesia as 1 mg/kg/hr of morphine. Higher infusion doses of both CMX-020 and morphi ne can be used to achieve much higher levels of analgesia.

[00103] s can be appreciated, CMX-020 is a fast-acting intravenous pain therapeutic that compa es in potency and efficacy to morphine. In animal tests, the most significant difference I etween CMX-020 and morphine is how fast the compound acts. With CMX- 020, the pei ik effect is reached within minutes; with morphine, the peak effect requires 15-40 minu tes. In continuous infusions, CMX-020 and morphine sustain similar analgesic k vels over 24 hours. While the required dose of CMX-020 is higher than morphine f >r continuous infusion, the added cost for a typical 6-10 mg CMX-020 dose for a 70 kg human would be a minimal.

[00104] The tail-flick and writhing assays represent different medical applications. The tail-flit k assay represents severe acute pain applications, such as acute trauma, intensive c< re, and break-through pain. For severe acute pain applications CMX-020 requires hi^ her doses. A prospective human dose for a 70 kg human for acute pain may be a 2.8 mg in a bolus injection for short-acting pain management, or 23.0 mg/hour in continuous infusions for longer term pain management. The writhing assay represents moderate p tin applications, including those associated with post-operative pain managemei it. CMX-020 may be administered at 0.6 mg in a bolus dose for short-acting application! or 6.0 mg/hour in continuous infusions for longer term pain management. A comparable dose of acetaminophen is 1000 mg.

33

QB\1 1803388. 128833.00( 10 PCT Application

Example 4 Exemplary AA analog formulations for analgesic treatment.

[00105] P reliminary tests show that CMX-020 is fully functional when delivered using the emulsic n vehicle formulation for Intralipid, as described in Table 5. Intralipid is used as an intrav snous nutritional supplement and has market precedent for delivery of therapeutic ; such as Propofol (marketed by AstraZeneca). CMX-020 can also be prepared w th emulsifying surfactants such as a lyophile, which is expected and typically used to pro 'ide long-term storage and then is diluted in standard vehicle for injection. Preparation of these two vehicles is described below in Table 5.

Table 5. Ingredients of Intralipid Formulation

Quantities: % (weight)

CMX-020 1%

j soy bean oi l 10%

egg phosphatide 1.20%

[glycerol 2.25%

i disodium edetate dihydrate 0.01%

sodium hydroxide (pH neutralizer)

water for injections to 100%

[00106] Preparation of Intralipid Vehicle. Variation #1: This variation has been demonstrat id successfully by the inventors with CMX-020 in animal models. The Intralipid fi irmulation is an oil-in-water emulsion. The ingredients for the Intralipid formulatiot that comprises 1 % of CMX-020 by weight is shown in Table 5. Briefly, the preparation is as follows: The aqueous phase, comprising glycerol, disodium edetate dehydrate, iodium hydroxide, and water are mixed and filtered. In parallel, an oil phase comprising soy bean oil, CMX-020, and egg phosphatide is stirred, filtered and added to the aqueou ; phase via a static mixer. The mixture is then circulated through a high pressure he mogenizer until a mean globule size of 250 nm is achieved. The emulsion is then filtere 1, filled into a container under nitrogen, and autoclaved. Variation #2: Since pure CMX- 020 is itself a light oil, the soy bean oil is not necessary and is not included. In this forn ulation variation, CMX-020 would simply replace soy bean oil as an ingredient. This variation would enable much higher therapeutic concentrations in the same emul: ion.

34

QB\1 1803388. 1 128833.000: 0 PCT Application

[00107] P reparation of Lyopbile Vehicle. The lyophile is an anhydrous emulsion composition that contains cryoprotection agents/bulking agents and can be redispersed by addition of \ 'ater and give the original water containing emulsion with nearly identical particle size distribution. The emulsion composition is prepared by removing the aqueous pha se by lyophilization, which is accomplished by freeze-drying.

Example 5: Exemplary AA analog pharmacokinetics.

[00108] F reliminary Results of LC/MS Pharmacokinetics of CMX-020. Mice were injected in ti e tail vein with 10 mg/kg of CMX-020. Venous blood samples were collected at 5, 15, 30 and 60 min following the injection of CMX-020. The blood samples wer ϊ centrifuged to collect plasma and CMX-020 was extracted from 15 uL of plasma with chloroform :methanol (2: 1 ). The extract was dried under a 2 stream at room temperature, resuspended in mobile phase and quantified by LC MS. As seen in Figure 6, the concentration of CMX-020 in blood plasma falls with a half-life of approximately 17 minutes. Extrapolation back to time zero represents an approximate plasma concentratio n of 14 ng/uL (or 33 uM). Samples of liver and brain were taken at 15 and 60 min after the single dose of CMX-020 was administered, homogenized and CMX-020 extracted as for plasma. The amount of CMX-020 present in brain was higher (704, 843 ng/g tissue, i=2) than that found in the liver (170, 21 1 ng/g tissue, n=2) at the 1 5 min timepoint. L ess than 20 ng/g tissue of CMX-020 was found in both brain and liver at the 60 min time x>int. These results suggest that there is a preferential distribution of CMX- 020 to the b ain which is thought to be the primary target organ for mediating the analgesic ef fect.

35

QB\1 1803388.1 128833.000 0 PCT Application

Example 6. Synthesis of AA analogs from Arachidonic Acid.

[00109] 1 he present example illustrates a method of synthesizing AA analogs according to the invention starting from arachidonic acid. The below method yields the preferred co npound CMX-020 but the method is suitable for providing related analogs through no r lore than routine optimization. (N, N-carbonyldiimidazole ("IM 2 CO"); Ν,Ν'- dicyclohexy carbodiimide ("DCC")

Example 7. De Novo Synthesis of AA analogs.

[00110] The present example illustrates the de novo synthesis of AA analogs of the invention. The illustrated synthesis yields the preferred analog CMX-020 but the method is applicable to the manufacture of related compounds, including the compounds shown in Figure 3, by no more than routine optimization.

SUMMARY OF CMX-020 DE NOVO SYNTHESIS

REACTIO> A

CsHgC 2 C 9 H 13 NO

Exact Mass: 112.05 Exact Mass: 151.10

ol. Wt.: 112.13 Mol. Wt.: 151.21

36

QB\1 1803388.1 128833.000 0 PCT Application

[00111] T o a stirred solution of 5-hexynoic acid (5 g, 44.59 mmol) in anhydrous CH2CI2 (50 ml) was added Ν,Ν'-dicyclohexylcarbodiimide ("DCC") (13.80 g, 66.88 mmol) folio wed by cyclopropylamine (2.80 g, 49.05 mmol) under an argon atmosphere at 0°C. Afte r 2 h at 0°C, TLC of the reaction mixture showed completion reaction. The white preci >itate was removed via filtration and the filtrate was concentrate under reduced pre ssure. The gummy residue was purified by SiC column chromatography using Biota; ;e pre-packed column (size: 100 g; solvent system: 10-75% EtOAc/hexane) to give pure amide 2 (6.54 g, 97%) as a white solid. Melting Point: 54.5-55.0°C.

C4H6 12 C11 H12O4S

Exact Mass : 86.04 Exact Mass: 240.05

Mol. Wt.: J6.09 Mol. Wt: 240.28

37

QB\ 1 1803388.1 128833.00010 PCT Application

[00112] r o a stirring 0°C solution of 2-butyn- l ,4-diol (10.00 g, 1 16.1 57 mmol) in anhydrous H 2 C1 2 (100 mL) was added anhydrous pyridine (18.276 g, 232.215 mmol) under an ari ;on atmosphere. TsCl (22.145 g, 1 16.157 mmol) was then added portionwise over a perio d of 15 min. After stirring for another l h at 0°C, TLC of the reaction mixture revealed an approx. 70:30 ratio of mono and di-tosylate, but no starting diol. Water was added to quench the reaction. The CH2CI2 layer was washed with water and CUSO4 solution, th< n finally dried over anhydrous Na2S0 4 . After evaporation of the solvent, the crude produ ct was purified by S1O2 column chromatography using a Biotage pre-packed column (siz ;: 340 g; solvent system: 10-50% EtOAc/hexane) to give pure mono-tosylate 3 (20.93 g, ' '5%) as a gummy semi-solid.

REACTION C

C 9 H 13 NO Γ ΐ1^12^4-5 C13H17NO2

Exact Mass : 151.10 Exact Mass: 240.05 Exact Mass: 219.13

Mol. Wt.: 11.1.21 Mol. Wt.: 240.28 Mol. Wt.: 219.28

[00113] 0 a stirring, heterogeneous mixture of acetylene 2 (1.0 g, 6.613 mmol), Cul (1.26 g, 6.έ 18 mmol), Nal (0.99 g, 6.618 mmol) and Cs 2 C0 3 (2.16 g, 6.613 mmol) in anhydrous ! DMF (50 mL) at 0°C was added mono-tosylate 3 (2.07 g, 8.603 mmol) in DMF (2 m ..) under an argon atmosphere. After stirring for 2 h at 0°C, the reaction mixture wai allow to warm slowly to room temperature and stirred for another 24 h. The reaction mi xture was diluted with ethyl acetate (200 mL) and a small amount of precipitate was removed via filtration. The filtrate was washed with water, brine and dried over 1 nhydrous Na 2 S0 4 . After evaporation of the solvent, the residue was purified

38

QB\1 1803388.1 128833.00010 PCT Application by S1O2 col jmn chromatography using a Biotage pre-packed column (size: 50 g; solvent system: 10- 100% EtOAc/hexane) to give pure di-acetylene 4 (1.17 g, 81%).

[00114] Di-acetylene 4 is very sensitive to auto-oxidation. Store under argon in a non- polar solve it like hexane or toluene that is oxygen free. Use in next step as soon as practical.

REACTION D

C, iH 17 N0 2 C 14 H, 9 N0 4 S

Ex act Mass: 219.13 Exact Mass: 297.10

Md. Wt: 219.28 Mol. Wt.: 297.37

[00115] " 0 a stirring, 0°C solution of alcohol 4 (1.17 g, 5.335 mmol) in anhydrous CH2CI2 (25 mL) was added Et 3 N (0.647 g, 6.402 mmol) under an argon atmosphere. MsCI (0.67: ! g, 5.869 mmol) was then added dropwise via syringe over 15 min. After 1 h at 0°C, the l eaction was quenched with water. The CH2CI2 layer was washed with water and brine, then dried over anhydrous Na 2 S0 4 . After evaporation of the solvent, the residue was purified by Si0 2 column chromatography using a Biotage pre-packed column (size: 100 g solvent system: 10-75% EtOAc/hexane) to give pure mesylate 5 (1.44 g, 91 %) as an >ff white solid. Melting Point: 88.0-88.5°C (decomposed)

REACTION E

39

QB\1 1803388. 1 128833.00010 PCT Application

MsCI, TEA

-OH "OMs

o°c

C 4 H 6 C C H8O3S

Exact Mass: 70.04 Exact Mass: 148.02

Mol. Wt.: 73.09 Mol. Wt.: 148.18

[00116] r , o a stirring, 0°C solution of 3-butyn-l-ol 6 (10.0 g, 142.67 mmol) in anhydrous CH2G2 (100 mL) was added TEA (21.65 g, 213.95 mmol) under an argon atmosphere MsCI (16.34 g, 1 1.04 mmol) was then added dropwise via syringe over a period of 1 f min. After 1 h at 0°C, the reaction was quenched with water. The CH2CI2 layer was ashed with water and brine, then dried over anhydrous Na2S0 4 . After evaporation of the solvent, the residue was purified by S1O2 column chromatography using a Bioi age pre-packed column (size: 340 g; solvent system: 0-30% EtOAc hexane) to give pure mesylate 7 (20.08 g, 95%).

REACTION F

C5H8O3S C 7 H 13 N

Exact Mass: 148.02 Exact Mass: 111.10 Mol. Wt: 148.18 Mol. Wt.: 111.18

40

QB\ 1 1803388. 128833.00010 PCT Application

[00117] Mesylate 7 (5.0 g, 33.74 mmol), isopropylamine (2.99 g, 50.61 mmol) and TEA (5.12 j ;, 7.05 mmol) were heated at 70°C in a sealed tube. After for 3 h, the reaction mixture wa; concentration and the residue was dried under high vacuum to crude N- isopropylbu t-3-yn-l -amine that was used in the next step without further purification.

REACTIO

.18

IV ol. Wt.: 1 11.18

[00118] To a stirring, 0°C solution of hexanoic acid (3.75 g, 33.72 mmol) in anhydrous < :H 2 C1 2 (50 mL) was added DCC ( 13.918 g, 67.44 mmol) followed by addition of DIPEA (17.43 g, 84.43 mmol) and the crude N-isopropylbut-3-yn-l -amine (3.75 g, 33. 72 mmol) from aboveunder an argon atmosphere. After stirring for 2 h at 0°C, the white pi ecipitate was removed via filtration. The filtrate was concentrate under reduced pre ssure, the gummy residue was purified by S1O2 column chromatography using a Biotage p ¾-packed column (size: 100 g; solvent system: 0-40% EtOAc/hexane) to give pure amide 8 (6.42 g) as a viscous oil.

REACTION H

41

QB\1 1803388. 128833.00010

Exact Mass: 410.29 Mol. Wt.: 410.59

[00119] " o a stirring, 0°C heterogeneous mixture of acetylene 8 (0.844 g, 4.035 mmol), Cul (0.768 g, 4.035 mmol), Nal (0.604 g, 4.035 mmol) and Cs 2 C0 3 (1.314 g, 4.035 mmo ) in anhydrous DMF (50 mL) was added a solution of mesylate 5 (1.2 g, 4.035 mmo ' ) in DMF (5 mL) under an argon atmosphere. After stirring for 2 h at 0°C, the reaction mi: :ture was slowly warmed to room temperature and stirred for an additional 24 h. The read ion mixture was then diluted with ethyl acetate (200 mL) the precipitated mass was re moved via filtration. The filtrate was washed with water, brine, dried over anhydrous Na 2 S0 4 , and evaporation in vacuo. The residue was purified by Si0 2 column chromatogr iphy using a Biotage pre-packed column (size: lOOg; solvent system: 10-75% EtOAc/hexi ne) to give /rw-acetylene 9 (1.3 g, 89%) as a pale yellow oil which was used in the next s tep as early as possible.

[00120] : "ra-acetylene 9 is extremely sensitive to auto-oxidation. Store under argon in a non-polar solvent like hexane or toluene that is oxygen free.

REACTION I

42

QB\1 1803388. 128833.00010 PCT Application

9 CMX-020

I3BN2O2 C26H44N2O2

Exa ;t Mass: 410.29 Exact Mass: 416.34 Mol. Wt.: 410.59 Mol. Wt.: 416.64

[00121] "0 a stirring, room temperature solution of nickel acetate tetra-hydrate (0.606 g, 2.435 mnol) in absolute EtOH (5 mL) was added solid NaBH 4 (0.092 g, 2.43 mmol) under a H 2 itmosphere (balloon ~ 1 atm). The resulting black suspension was stirred for 30 min, the n distilled ethylenediamine (0.658 g, 10.948 mmol) was added via syringe. Following complete addition, the suspension was stirred for another 15 min, then triacetylene 9 (0.50 g, 1.217 mmol) in absolute EtOH (5 mL) was added. After stirring at room temperature under H 2 (balloon ~ 1 atm) for 3 h, TLC showed completion of the reaction. D ethyl ether (50 mL) was added to dilute the reaction mixture that was then passed thro agh a short silica gel column to remove catalyst and ethylenediamine. The filtrate was concentrated under reduced pressure and concentrated in vacuo. The residue was purifie d by flash column chromatography (0-75% ethyl acetate/hexanes) to give CMX-020 is a colorless oil (91 %). HPLC analysis (CI 8, 70/30 acetonitrile H 2 0) of this product she ed about 10% of over saturated product, which was further purified by prep HPLC to g .ve CMX-202 as a 99% pure product as a viscous oil. CMX-020 is stored under argor at -20°C or -80°C.

Example 8 Antipyretic Effect of CMX-020

[00122] This example demonstrates the antipyretic effect of compound CMX-020 as compared t > morphine. Recordings of body temperature in male mice (30 grams) were

43

QB\1 1803388. 128833.00010 PCT Application achieved us ing a mouse rectal temperature probe. Baseline was recorded twice for 10 minutes at ntervals of 2 minutes with a 15 minute span of no recording. After an i.v. bolus inject on of either CMX-020 (l Omg/kg) or morphine (10 mg/kg) body temperature was recorde d every 2 minutes for 50 minutes. Figure 7 illustrates the antipyretic effect of compound < ^MX-020 as compared to morphine.

[00123] I )ther embodiments and uses of the invention will be apparent to those skilled in the art from consideration from the specification and practice of the invention disclosed h< rein. All references cited herein for any reason, including all journal citations an i U.S./foreign patents and patent applications, are specifically and entirely incorporated herein by reference. It is understood that the invention is not confined to the specific rea jents, formulations, reaction conditions, etc., herein illustrated and described, but embract s such modified forms thereof as come within the scope of the following claims.

44

QB\1 1803388.