WO2011143521A2

WO2011143521A2 - Enhanced homing and engraftment of hematopoietic stem cells using cd26 inhibitors

Info

Publication number: WO2011143521A2
Application number: PCT/US2011/036380
Authority: WO
Inventors: Diana Severynse-Stevens; John R. Didsbury
Original assignee: Dara Biosciences, Inc.
Priority date: 2010-05-13
Filing date: 2011-05-13
Publication date: 2011-11-17
Also published as: WO2011143521A3

Abstract

The present invention relates to methods for improving the therapeutic usefulness of hematopoietic stem cells (HSC). Specifically, the invention involves contacting HSC with compounds of the invention to enhance homing and/or engraftment to bone marrow and HSC thus contacted.

Description

Enhanced Homing and Engraftment of Hematopoietic Stem Cells

Using CD26 Inhibitors

FIELD OF THE INVENTION

[0001] The present invention relates to methods for improving the therapeutic usefulness of hematopoietic stem cells (HSC). Specifically, the invention involves contacting HSC with compounds of the invention to enhance homing and/or engraftment to bone marrow as well as HSC thus contacted.

BACKGROUND OF THE INVENTION

[0002] Stem cell transplants, in particular bone marrow transplants using HSC, are an important procedure for therapy of hematological disorders, including leukemia, lymphoma, myelodysplasia, and anemia. However, the efficiency of bone marrow transplantation can be very low, requiring the transplantation of large numbers of stem cells and a long time period for full immune system and platelet recovery. Overall patient survival for bone marrow transplants is as low as 50%.

[0003] HSC for transplantation are traditionally provided by matching bone marrow donors, but thousands of patients die each year waiting for a matching donor to be identified. Umbilical cord blood is an increasingly important source for HSC. However, the low number of HSC in a single unit of umbilical cord blood is insufficient for use in adults.

[0004] A need exists for a more efficient HSC transplantation method exhibiting one or more advantages, including requiring fewer cells per transplant, accelerating immune system and platelet recovery, and/or improving patient survival outcomes.

SUMMARY OF THE INVENTION

[0005] The present invention is based, in part, on the ability of CD26 (also known as dipeptidylpeptidase IV (DPPIV)) inhibitors to enhance the homing and/or engraftment of HSC. Accordingly, as one aspect, the invention provides methods for increasing homing and/or engraftment of HSC to bone marrow comprising contacting HSC with an effective amount of a compound of Formula I:

wherein:

X is NR³ or O;

n is 1 or 2;

A is a bicyclic or tricyclic carbocycle of 5 to 20 atoms wherein each bridge of the bicycle has at least one atom;

wherein:

p and q are independently 0 or 1 ;

Y is CH₂, CHF, CF₂, O, or S(0)_m;

W and Z are independently CH₂, CHF, or CF₂;

and wherein the ring formed by N, W, Y, Z and the carbon atoms to which they are attached is saturated or optionally contains one double bond;

when X is NR³ then R² is R⁴-S0₂-; R⁵-S0₂- H-C(0)-; R⁶R⁷N-S0₂-; or a heterocyclic group unsubstituted or optionally substituted with halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, haloalkyloxy, cycloalkyloxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocycloalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, cycloalkyl-S(0)_m, cycloalkylalkyl-S(0)_m, aryl-S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano; or

when X is O then R² is a heterocyclic group unsubstituted or optionally substituted with halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, haloalkyloxy, cycloalkyloxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocycloalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, cycloalkyl-S(0)_m, cycloalkylalkyl-S(0)_m, aryl- S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano;

R³ is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl;

R⁴ is selected from the group consisting of haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl;

R⁵ is selected from the group consisting of alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl;

R⁶ and R⁷ are each independently selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl; or R⁶ and R⁷ together form C₃-C₇ alkylene;

R⁸ is H or cyano;

m is 0, 1 or 2;

or a pharmaceutically acceptable salt or prodrug thereof.

[0006] In certain embodiments, the HSC are contacted with the compounds of the invention in vitro, ex vivo, or in vivo. The HSC can further be fucosylated. The contacted HSC can then be delivered to a subject in need of a transplant, e.g., a bone marrow transplant.

[0007] A further aspect of the invention relates to compositions comprising the contacted HSC of the invention.

[0008] Another aspect of the invention relates to methods for treating a condition in a subject for which transplantation of HSC is required or advantageous, comprising providing a composition of HSC of the invention, and delivering to the subject a therapeutically effective quantity of the composition.

[0009] An additional aspect of the invention relates to methods for performing a bone marrow transplantation in a subject, comprising providing a composition of HSC of the invention, and delivering to the subject a therapeutically effective quantity of the composition.

[0010] A further aspect of the invention relates to kits comprising the HSC and/or compounds of the invention.

DETAILED DESCRIPTION OF THE INVENTION

[0011] The present invention will now be described in more detail, in which preferred embodiments of the invention are shown. This invention may, however, be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

[0012] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. All publications, patent applications, patents, patent publications and other references cited herein are incorporated by reference in their entireties for the teachings relevant to the sentence and/or paragraph in which the reference is presented.

[0013] Unless the context indicates otherwise, it is specifically intended that the various features of the invention described herein can be used in any combination. Moreover, the present invention also contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted. To illustrate, if the specification states that a complex comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, can be omitted and disclaimed.

I. Definitions

[0014] As used in the description of the invention and the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0015] Also as used herein, "and/or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or").

[0016] The term "about," as used herein when referring to a measurable value such as an amount of compound, dose, time, temperature, enzymatic activity or other biological activity and the like, is meant to encompass variations of ± 20%, ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of the specified amount.

[0017] As used herein, the transitional phrase "consisting essentially of is to be interpreted as encompassing the recited materials or steps "and those that do not materially affect the basic and novel characteristic(s)" of the claimed invention (e.g., enhanced homing and/or engraftment). See, In re Herz, 537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463 (CCPA 1976) (emphasis in the original); see also MPEP § 2111.03. Thus, the term "consisting essentially of as used herein should not be interpreted as equivalent to "comprising." [0018] The term "halo," as used herein, refers to any suitable halogen, including -F, - CI, -Br, and -I.

[0019] The term "mercapto," as used herein, refers to an -SH group.

[0020] The term "azido," as used herein, refers to an -N₃ group.

[0021] The term "cyano," as used herein, refers to a -CN group.

[0022] The term "hydroxyl," as used herein, refers to an -OH group.

[0023] The term "nitro," as used herein, refers to an -N0₂ group.

[0024] The term "alkyl," as used herein alone or as part of another group, refers to a straight or branched chain hydrocarbon containing from 1 to 10 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n- hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, and the like. "Lower alkyl," as used herein, is a subset of alkyl, and refers to a straight or branched chain hydrocarbon group containing from 1 to 4 carbon atoms. Representative examples of lower alkyl include, but are not limited to, methyl, ethyl, n- propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, and the like. The term "alkyl" or "lower alkyl" is intended to include both substituted and unsubstituted alkyl or lower alkyl unless otherwise indicated and these groups may be substituted with groups selected from halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, alkenyloxy, alkynyloxy, haloalkoxy, cycloalkoxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocyclolalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, alkenyl-S(0)_m, alkynyl-S(0)_m, cycloalkyl- S(0)_m, cycloalkylalkyl-S(0)_m, aryl-S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, alkenylamino, alkynylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro, or cyano where m= 0, 1 or 2.

[0025] The term "alkenyl," as used herein alone or as part of another group, refers to a straight or branched chain hydrocarbon containing from 1 to 10 carbon atoms (or in lower alkenyl 1 to 4 carbon atoms) which include 1 to 4 double bonds in the normal chain. Representative examples of alkenyl include, but are not limited to, vinyl, 2-propenyl, 3- butenyl, 2-butenyl, 4-pentyl, 3-pentyl, 2-hexenyl, 3-hexenyl, 2,4-heptadiene, and the like. The term "alkenyl" or "lower alkenyl" is intended to include both substituted and unsubstituted alkenyl or lower alkenyl unless otherwise indicated and these groups may be substituted with groups as described in connection with alkyl and lower alkyl above.

[0026] The term "alkynyl," as used herein alone or as part of another group, refers to a straight or branched chain hydrocarbon containing from 1 to 10 carbon atoms (or in lower alkynyl 1 to 4 carbon atoms) which include 1 triple bond in the normal chain. Representative examples of alkynyl include, but are not limited to, 2-propynyl, 3-butynyl, 2-butynyl, 4-pentenyl, 3-pentenyl, and the like. The term "alkynyl" or "lower alkynyl" is intended to include both substituted and unsubstituted alkynyl or lower alkynyl unless otherwise indicated and these groups may be substituted with the same groups as set forth in connection with alkyl and lower alkyl above.

[0027] The term "alkoxy," as used herein alone or as part of another group, refers to an alkyl or lower alkyl group, as defined herein, appended to the parent molecular moiety through an oxy group, -0-. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, tert-butoxy, pentyloxy, hexyloxy and the like.

[0028] The term "acyl," as used herein alone or as part of another group, refers to a -C(0)R radical, where R is any suitable substituent such as aryl, alkyl, alkenyl, alkynyl, cycloalkyl or other suitable substituent as described herein.

[0029] The term "haloalkyl," as used herein alone or as part of another group, refers to at least one halogen, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of haloalkyl include, but are not limited to, chloromethyl, 2-fluoroethyl, trifluoromethyl, pentafluoroethyl, 2-chloro-3- fluoropentyl, and the like.

[0030] The term "alkylthio," as used herein alone or as part of another group, refers to an alkyl group, as defined herein, appended to the parent molecular moiety through a thio moiety, as defined herein. Representative examples of alkylthio include, but are not limited to, methylthio, ethylthio, tert-butylthio, hexylthio, and the like.

[0031] The term "aryl," as used herein alone or as part of another group, refers to a monocyclic carbocyclic ring system or a bicyclic carbocyclic fused ring system having one or more aromatic rings. Representative examples of aryl include, but are not limited to, azulenyl, indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl, and the like. The term "aryl" is intended to include both substituted and unsubstituted aryl unless otherwise indicated and these groups may be substituted with the same groups as set forth in connection with alkyl and lower alkyl above. [0032] The term "arylalkyl," as used herein alone or as part of another group, refers to an aryl group, as defined herein, appended to the parent molecular moiety through an alkyl group, as defined herein. Representative examples of arylalkyl include, but are not limited to, benzyl, 2-phenylethyl, 3-phenylpropyl, 2-naphth-2-ylethyl, and the like.

[0033] The term "amino," as used herein alone, means the radical -NH₂.

[0034] The term "alkylamino," as used herein alone or as part of another group, means the radical -NHR, where R is an alkyl group.

[0035] The term "arylalkylamino," as used herein alone or as part of another group, means the radical -NHR, where R is an arylalkyl group.

[0036] The term "disubstituted-amino," as used herein alone or as part of another group, means the radical -NR_aR_b, where Ra and R_b are independently selected from the groups alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl.

[0037] The term "acylamino," as used herein alone or as part of another group, means the radical -NR_aR_b, where R_a is an acyl group as defined herein and R_b is selected from the groups hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl.

[0038] The term "acyloxy," as used herein alone or as part of another group, means the radical -OR, where R is an acyl group as defined herein.

[0039] The term "ester," as used herein alone or as part of another group, refers to a - C(0)OR radical, where R is any suitable substituent such as alkyl, cycloalkyl, alkenyl, alkynyl, or aryl.

[0040] The term "amide," as used herein alone or as part of another group, refers to a - C(0)NRaR_b radical, where Ra and R_b are any suitable substituent such as alkyl, cycloalkyl, alkenyl, alkynyl, or aryl.

[0041] The term "sulfonamide," as used herein alone or as part of another group, refers to a -S(0)₂NRaR_b radical, where Rg and R_b are any suitable substituent such as H, alkyl, cycloalkyl, alkenyl, alkynyl, or aryl.

[0042] The term "urea," as used herein alone or as part of another group, refers to an - N(Rc)C(0)NRaR_b radical, where Ra, R_b, and R_c are any suitable substituent such as H, alkyl, cycloalkyl, alkenyl, alkynyl, or aryl.

[0043] The term "alkoxyacylamino," as used herein alone or as part of another group, refers to an -N(R_a)C(0)OR_b radical, where R_a and R_b are any suitable substituent such as H, alkyl, cycloalkyl, alkenyl, alkynyl, or aryl. [0044] The term "aminoacyloxy," as used herein alone or as part of another group, refers to an -OC(0)NRaR_b radical, where R_a and R_b are any suitable substituent such as H, alkyl, cycloalkyl, alkenyl, alkynyl, or aryl.

[0045] The term "cycloalkyl," as used herein alone or as part of another group, refers to a saturated. or partially unsaturated cyclic hydrocarbon group containing 3, 4, 5, 6, 7, or 8 carbons. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. These rings may be optionally substituted with halo or lower alkyl.

[0046] The terms "heterocyclic group" or "heterocyclo," as used herein alone or as part of another group, refer to a monocyclic or a bicyclic ring system. Monocyclic ring systems are exemplified by any 5, 6, 7, or 8 membered ring containing 1 , 2, 3, or 4 heteroatoms independently selected from oxygen, nitrogen and sulfur. The 5 membered ring has from 0-2 double bonds and the 6 membered ring has from 0-3 double bonds. Representative examples of monocyclic ring systems include, but are not limited to, azetidine, azepine, aziridine, diazepine, 1,3-dioxolane, dioxane, dithiane, furan, imidazole, imidazoline, imidazolidine, isothiazole, isothiazoline, isothiazolidine, isoxazole, isoxazoline, isoxazolidine, morpholine, oxadiazole, oxadiazoline, oxadiazolidine, oxazole, oxazoline, oxazolidine, piperazine, piperidine, pyran, pyrazine, pyrazole, pyrazoline, pyrazolidine, pyridine, pyrimidine, pyridazine, pyrrole, pyrroline, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, tetrazine, tetrazole, thiadiazole, thiadiazoline, thiadiazolidine, thiazole, thiazoline, thiazolidine, thiophene, thiomorpholine, thiomorpholine sulfone, thiopyran, triazine, triazole, trithiane, and the like. Bicyclic ring systems are exemplified by any of the above monocyclic ring systems fused to an aryl group as defined herein, a cycloalkyl group as defined herein, or another monocyclic ring system as defined herein. Representative examples of bicyclic ring systems include, but are not limited to, for example, benzimidazole, benzothiazole, benzothiadiazole, benzothiophene, benzoxadiazole, benzoxazole, benzofuran, benzopyran, benzothiopyran, benzodioxine, 1 ,3-benzodioxole, cinnoline, indazole, indole, indoline, indolizine, naphthyridine, isobenzofuran, isobenzothiophene, isoindole, isoindoline, isoquinoline, phthalazine, purine, pyranopyridine, quinoline, quinolizine, quinoxaline, quinazoline, tetrahydroisoquinoline, tetrahydroquinoline, thiopyranopyridine, and the like. These rings may be optionally substituted with groups selected from halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, alkenyloxy, alkynyloxy, haloalkoxy, cycloalkoxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocyclolalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, alkenyl-S(0)_m, alkynyl- S(0)_m, cycloalkyl-S(0)_m, cycloalkylalkyl-S(0)_m, aryl-S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, alkenylamino, alkynylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano where m = 0, 1 or 2.

[0047] The terms "treat," "treating," and "treatment," as used herein, refer to any type of treatment that imparts a benefit to a patient afflicted with a disease, including improvement in the condition of the patient (e.g., in one or more symptoms), delay in the progression of the disease, etc.

[0048] The term "pharmaceutically acceptable," as used herein, means that the compound or composition is suitable for administration to a subject to achieve the treatments described herein, without unduly deleterious side effects in light of the severity of the disease and necessity of the treatment.

[0049] The term "physiologically acceptable," as used herein, means that the compound or composition is suitable for administration to a subject, without unduly deleterious side effects.

[0050] The term "pharmaceutically acceptable prodrugs," as used herein, refers to those prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and other animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable risk/benefit ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention. The term "prodrug" refers to compounds that are transformed in vivo to yield the parent compound of the above formulae, for example, by hydrolysis in blood. A thorough discussion is provided in T. Higuchi and V. Stella, Prodrugs as Novel delivery Systems, Vol. 14 of the A.C.S. Symposium Series and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated by reference herein. See also U.S. Patent No. 6,680,299 Examples include a prodrug that is metabolized in vivo by a subject to an active drug having an activity of compounds as described herein, wherein the prodrug is an ester of an alcohol or carboxylic acid group, if such a group is present in the compound; an acetal or ketal of an alcohol group, if such a group is present in the compound; an N-Mannich base or an imine of an amine group, if such a group is present in the compound; or a Schiff base, oxime, acetal, enol ester, oxazolidine, or thiazolidine of a carbonyl group, if such a group is present in the compound, such as described in U.S. Patent No. 6,680,324 and U.S. Patent No. 6,680,322. Prodrugs of the present invention include esters or compositions as described in U.S. Patent No. 6,548,668 to Adams et al, U.S. Patent No. 6,083,903 to Adams et al, or U.S. Patent No. 6,699,835 to Plamondon et al, the disclosures of which are incorporated by reference herein in their entirety.

[0051] "CD26" (DPPIV/dipeptidylpeptidase IV) is a membrane bound extracellular peptidase that cleaves dipeptides from the N-terminus of polypeptide chains after a proline or an alanine. "CD26 activity" or "DPPIV activity" encompasses any activity of CD26, including peptidase activity. "CD26" or "DPPIV" inhibitor or antagonist refers to any substance, chemical, biological, and so forth, which is capable of inhibiting CD26 activity. Preferably, CD26 or DPPIV inhibitors and antagonists inhibit CD26 peptidase activity, at levels sufficient to improve stem cell homing. In representative embodiments, the inhibitor inhibits CD26 activity by at least about 20%, e.g., about 30, 40, 50, 60, 70, 80, or 90% or more. CD26 activity can be routinely measured by assays well known in the art.

[0052] The term "homing," as used herein, refers to localization to a particular area, for example localization of transplanted stem cells to the bone marrow.

[0053] The terms "engrafting" or "engraftment," as used herein, means the persistence of proliferating stem cells in a particular location over time. The stem cells can persist, e.g., for at least about 1 day, e.g., at least about 2, 3, 4, 5, or 6 days or at least about 1, 2,

3, 4, 5, 6, 7, or 8 weeks or more, or even one or more months or years.

[0054] The term "effective amount," as used herein in reference to the compounds of the invention, refers to an amount of the compounds of the invention that is sufficient to alter HSC to increase their ability to home and/or engraft in bone marrow.

[0055] The term "donor," as used herein, refers to the organism donating the HSC.

[0056] The term "recipient," as used herein, is the subject receiving the HSC.

[0057] The terms "stem cell" or "hematopoietic stem cell," as used herein, means a pluripotent cell of the hematopoietic system capable of differentiating into a cell of a specific lineage, such as lymphoid or myeloid.

[0058] The term "myelosuppressed," as used herein, refers to a subject {e.g., a patient) who has undergone irradiation, or other treatment, such as chemotherapeutic treatment, to cause the death of at least 10% of the bone marrow cells of the subject. [0059] The term "myeloablated," as used herein, refers to a subject (e.g., a patient) who has undergone irradiation, or other treatment, such as chemotherapeutic treatment, to cause the death of at least 90% of the bone marrow cells of the subject.

[0060] The term "non-myeloablated," as used herein, refers to a subject who has not undergone irradiation, or other treatment (such as chemotherapy) to cause the death of the bone marrow cells of the subject.

[0061] The term "autologous," as used herein, refers to transfer of cells involving one subject as both donor and recipient.

[0062] The term "syngeneic," as used herein, refers to donor cells that are genetically identical or closely related to the cells of a subject, so as to allow tissue transplantation.

[0063] The term "allogeneic," as used herein, refers to donor cells that are genetically different from the cells of a subject although belonging to or obtained from the same species.

[0064] The term "pluripotent," as used herein, means that a cell is capable, through its progeny, of giving rise to all of the cell types which comprise the adult animal including the germ cells. Both embryonic stem and embryonic germ cells are pluripotent cells under this definition.

II. Compounds

[0065] One aspect of the present invention relates to methods for increasing homing and/or engraftment of HSC to bone marrow using compounds of the invention. The compounds of the present invention are described in International Publication No. WO 2006/012395 (incorporated herein by reference in its entirety) and are illustrated by Formula I:

wherein:

X is NR³ or O;

n is 1 or 2;

wherein:

p and q are independently 0 or 1 ;

Y is CH₂, CHF, CF₂, O, or S(0)_m;

W and Z are independently CH₂, CHF, or CF₂;

when X is NR³ then R² is R⁴-S0₂-; R⁵-S0₂-NH-C(0)-; R⁶R⁷N-S0₂-; or a heterocyclic group unsubstituted or optionally substituted with halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, haloalkyloxy, cycloalkyloxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocycloalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, cycloalkyl-S(0)_m, cycloalkylalkyl-S(0)_m, aryl-S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano; or

R⁴ is selected from the group consisting of haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl; R⁵ is selected from the group consisting of alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl;

R⁶ and R⁷ are each independently selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl; or R⁶ and R⁷ together form C3-C7 alkylene;

R⁸ is H or cyano;

m is 0, 1 or 2;

or a pharmaceutically acceptable salt or prodrug thereof.

[0066] Examples of suitable groups "A" include, but are not limited to, adamantyl, bicyclo[2.1 , l]hexane, bicyclo[3.1.1]heptane, bicyclo[3.2.1]octane, bicyclo[2.2.2]octane, and bicyclo[3.3.1]nonane, which may optionally include one or more double bonds.

[0067] Examples of suitable adamantyl groups "A" for carrying out the present invention, with linkages, include, but are not limited to, the following:

[0068] Particular examples of suitable groups "A" for carrying out the present invention, with linkages, include, but are not limited to, the following:

(a bicyclo[2.2.2]octane) (a bicyclo[3.2.1]octane) (a bicyclo[3.1.1]heptane)

[0069] In one embodiment, X is NR³ and R² is R⁴-S0₂-, R⁵-S0₂- H-C(0)-, R⁶R⁷N- S0₂- or a heterocyclic group. In another embodiment, X is O and R² is a heterocyclic group

[0070] In one embodiment, the compound is:

or a pharmaceutically acceptable salt or prodrug thereof.

[0071] In another embodiment, the compound is:

or a pharmaceutically acceptable salt or prodrug thereof.

[0072] In one embodiment, the compound is

or a pharmaceutically acceptable salt or prodrug thereof.

[0073] In one embodiment, Y is selected from the group consisting of CHF, CF₂, O, and S(0)_m; or q is 1 and W is selected from the group consisting of CHF and CF₂; or p is 1 and Z is selected from the group consisting of CHF and CF₂. In a further embodiment, Y is selected from the group consisting of CHF, CF₂, O, and S(0)_m; q is 1 and W is CH₂; and p is 0. In another embodiment, Y is selected from the group consisting of CHF, CF₂, O, and S(0)_m; or q is 0; and p is 1 and Z is CH₂. In a further embodiment, Y is CH₂; q is 1 and W is selected from the group consisting of CHF and CF₂; and p is 0. In a further embodiment, Y is CH₂; q is 0; and p is 1 and Z is selected from the group consisting of CHF and CF₂.

[0074] Examples of compounds of the present invention include, but are not limited to:

and pharmaceutically acceptable salts and prodrugs thereof.

[0075] Additional examples of compounds of the invention include, but are not limited to:

- 16 -

- 17-

- 18-

- 19-

-20-

-21 -

and pharmaceutically acceptable salts and prodrugs thereof.

[0076] In one embodiment, the compound is selected from the group consisting of

and pharmaceutically acceptable salts and prodrugs thereof.

[0077] Certain of the compounds described herein contain one or more chiral centers, or may otherwise be capable of existing as multiple stereoisomers. The scope of the present invention includes pure stereoisomers as well as mixtures of stereoisomers, such as purified enantiomers/diastereomers, enantiomerically/diastereomerically enriched mixtures, or racemates.

[0078] The compounds disclosed herein can, as noted above, be prepared in the form of their pharmaceutically acceptable salts. Pharmaceutically acceptable salts are salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects. Examples of such salts are (a) acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; and salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p- toluenesulfonic acid, naphthalenedisulfomc acid, polygalacturonic acid, and the like; (b) salts formed from elemental anions such as chlorine, bromine, and iodine, and (c) salts derived from bases, such as ammonium salts, alkali metal salts such as those of sodium and potassium, alkaline earth metal salts such as those of calcium and magnesium, and salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine.

[0079] Compounds of the present invention (this term including pharmaceutically acceptable salts and prodrugs thereof) can be made in accordance with known techniques {see, e.g., U.S. Patent No. 6,166,063 to Villhauer et al. and International Publication No. WO 2006/012395) or variations thereof which will be apparent to those skilled in the art based on the disclosure provided herein.

III. Methods

[0080] The present invention is based, in part, on the ability of CD26 inhibitors to enhance the homing and/or engraftment of HSC. Accordingly, as one aspect, the invention provides methods for increasing homing and/or engraftment of HSC to bone marrow, comprising contacting HSC with an effective amount of a compound of Formula I or a pharmaceutically acceptable salt or prodrug thereof as disclosed above. The methods can be carried out in vitro, ex vivo, or in vivo, or any combination thereof.

[0081] In in vitro embodiments, the HSC are obtained from an in vitro source, e.g., cell culture, and the HSC are contacted with a compound of the invention in vitro.

[0082] In ex vivo embodiments, the HSC are obtained from a subject, e.g., a human, e.g., from peripheral blood, umbilical cord blood, or bone marrow, and the HSC are contacted outside the body of the subject. Ex vivo embodiments include obtaining HSC from a subject and culturing the cells for a period of time prior to use (e.g., for transplantation). In some embodiments, after contact the cells are delivered to a subject, e.g., the same subject from which the cells were isolated (autologous donation) or a different subject (non-autologous (e.g., syngeneic or allogeneic) donation).

[0083] For in vivo embodiments, the HSC are contacted with a compound of the invention in the body of a subject. In some embodiments, the HSC to be contacted inside the body were delivered to the subject and the compound is separately delivered to the subject. The compound can be delivered to the subject before, during, and/or after administration of HSC to the subject. For example, the compound can be delivered about 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 12, 18, or 24 hours or more before and/or after delivery of HSC to the subject, as long as the delivery of the compound is effective to increase the homing and/or engraftment of the HSC. In other embodiments, the HSC and the compound are delivered together to a subject, e.g., as part of the same composition or separate compositions.

[0084] In certain embodiments, the in vitro, ex vivo, and/or in vivo methods can be combined. For example, the HSC can be contacted with the compound ex vivo and then delivered to a subject. More of the same compound and/or a different compound can be delivered to the same subject before, during and/or after delivery of the HSC.

[0085] The contacting step is carried out for a time sufficient for the HSC to exhibit increased homing and/or engraftment. The length of time will depend on the compound and the cells and can be readily determined using techniques well known in the art, such as those disclosed in U.S. Published Application No. 2005/0247574 and U.S. Patent No. 7,332,334. In certain embodiments, the length of the contact step can be about 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours or more.

[0086] In in vitro and ex vivo embodiments, the contacting step is carried out in a suitable container with a concentration of the compound of the invention sufficient to increase the homing and/or engraftment of the HSC. Suitable concentrations will depend on the compound and the cells, will depend on the length of time of the contacting step, and can be readily determined using techniques well known in the art, such as those disclosed in U.S. Published Application No. 2005/0247574 and U.S. Patent No. 7,332,334. In certain embodiments, the concentration of the compound can be about 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, or 100 mM or more. The concentration of cells in the container can be any concentration suitable for increasing homing and/or engraftment, e.g., about 1 x 10⁵ to about 1 x 10⁷ cells/ml, e.g., about 1 x 10⁶ cells/ml.

[0087] For in vivo embodiments, the contacting step involves delivering the compound to a subject in an amount and time sufficient to increase the homing and/or engraftment. The dose of compound delivered to a subject will depend on factors such as the age, gender, and health of the subject and the compound being delivered and can be readily determined by a health care practitioner. In certain embodiments, the dose of compound can be about 0.1 to about 100 μιηοΐ/kg body weight, e.g., about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 μιηοΐ/kg body weight or any range therein.

[0088] In some embodiments, the HSC are mammalian cells, e.g., human cells. Stem cells may be obtained by various techniques well known in the art. For example, cells may be from an autologous donor (the patient who will receive the cells), or from a non- autologous donor. Stem cells may be harvested from the bone marrow, obtained from umbilical cord blood, or isolated from peripheral blood cells (e.g., following G-CSF mobilizing agent treatment).

[0089] The HSC can be used as part of a mixed population of cells or partially or completely purified from other cells present in the source. In some embodiments, HSC can be contacted with the compounds of the invention before separation of the desired HSC from the unfractionated blood or bone marrow sample, for example, using total mononuclear cells from cord blood, peripheral blood, or bone marrow. In other embodiments, HSC are contacted after they are isolated.

[0090] Various techniques may be employed to separately obtain the desired population of HSC, e.g., CD34⁺CD38^low/" stem cells alone, or in combination with CD34⁺CD38⁺ HSC. Monoclonal antibodies are particularly useful for identifying markers (surface membrane proteins) associated with particular cell lineages and/or stages of differentiation. The antibodies may be attached to a solid support to allow for crude separation. The separation techniques employed can be selected to maximize the retention of viability of the fraction to be collected. The particular technique employed will depend upon efficiency of separation, cytotoxicity of the methodology, ease and speed of performance, and necessity for sophisticated equipment and/or technical skill.

[0091] Procedures for separation may include magnetic separation, using antibody- coated magnetic beads, and "panning" with antibody attached to a solid matrix, e.g., plate, or other convenient technique. Techniques providing accurate separation include fluorescence activated cell sorters, which can have varying degrees of sophistication, e.g., a plurality of color channels, low angle and obtuse light scattering detecting channels, and impedance channels.

[0092] Conveniently, the antibodies may be conjugated with markers, such as magnetic beads, which allow for direct separation; biotin, which can be removed with avidin or streptavidin bound to a support; fluorochromes, which can be used with a fluorescence activated cell sorter (FACS), or the like, to allow for ease of separation of the particular cell type. Any technique may be employed which is not unduly detrimental to the viability of the remaining cells.

[0093] In one embodiment, HSC lacking mature cell markers may be substantially enriched, wherein the cells may then be separated by the FACS or other methodology having high specificity. Multi-color analyses may be employed with the FACS which is particularly convenient. The cells may be separated on the basis of the level of staining for the particular antigens. Fluorochromes, which may find use in a multi-color analysis, include phycobiliproteins, e.g., phycoerythrin and allophycocyanins, fluorescein, and Texas red, for example.

[0094] In certain embodiments of the invention, in addition to being contacted by a compound of the invention, HSC can be fucosylated to increase their homing ability. HSC that have been fucosylated have enhanced binding to P-selectin and E-selectin, thereby promoting homing ability. Fucosylation can occur by contacting the HSC in vitro or ex vivo with an al,3 fucosyltransferase and a fucose donor. Fucosylation can be carried out before, during, and/or after contacting the HSC with the compounds of the invention. The al,3 fucosyltransferase may be, for example, an al,3 fucosyltransferase IV, an al,3 fucosyltransferase VI, or an al,3 fucosyltransferase VII, or a combination thereof. The fucose donor may be, for example, GDP -fucose.

[0095] In one embodiment, the HSC may be treated by adding free fucosyltransferase to the cell composition, wherein the final product containing the fucosylated cells also contains the fucosyltransferase which was used to treat the cells. In another embodiment, the HSC may be treated using fucosyltransferases which are bound to a support, such as magnetic beads, or any other support known by those of skill in the art, which can be separated from the cell composition after the treatment process is complete. Techniques for fucosylating HSC are described in U.S. Patent No. 7,332,334, U.S. Published Application No. 2006/0228340, and International Publication No. WO 2004/094619, each incorporated herein by reference in its entirety.

[0096] Another aspect of the invention relates to methods for treating a condition in a subject for which transplantation of HSC is required or advantageous, comprising providing a composition of HSC produced by the methods of the invention, and delivering to the subject a therapeutically effective quantity of the composition. Any condition in which a transplantation of HSC is required or advantageous may be treated by this method. Examples of such conditions include, without limitation, leukemias and lymphomas such as chronic myelocytic (myelogenous) leukemia (CML), juvenile chronic myelogenous leukemia (JCML), chronic lymphocytic leukemia, acute myelocytic leukemia (AML), acute lymphocytic leukemia (ALL), malignant lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, multiple myeloma, aplastic anemia gravis, sickle cell anemia, myelodysplastic syndrome (MDS), and autoimmune diseases, for example.

[0097] Other diseases that may be treated with the HSC of the present invention are Gunther's disease, Hunter syndrome, Hurler syndrome, neuroblastoma, Wiskott-Aldrich syndrome, X-linked lympho-proliferative syndrome, and solid tissue tumors, such as breast cancer. [0098] A further aspect of the invention relates to methods for performing a bone marrow transplantation in a subject, comprising providing a composition of HSC produced by the methods of the invention, and delivering to the subject a therapeutically effective quantity of the composition. In one embodiment, the cells can be used to reconstitute the bone marrow of a myelosuppressed or myeloablated subject, e.g., one that has been irradiated or subjected to chemotherapy.

[0099] The transplantation methods can comprise a further step of delivering a compound of the invention to the subject, before, during, and/or after delivery of the HSC.

[0100] Methods of HSC transplantation and bone marrow transplantation are well known in the art and can be readily carried out using the cells and compositions of the present invention. The cells of the present invention can be delivered by intravenous injection, for example, or by any other appropriate method known by those of skill in the art. In methods for treating a subject afflicted with a disease or condition, a

therapeutically effective amount of HSC is that amount sufficient to treat (e.g., reduce or eliminate) the symptoms or effects of the disease or condition. The therapeutically effective amount administered to a subject will be determined on an individual basis and will be based, at least in part, on consideration of the subject's size, the severity of symptoms to be treated, and the results sought. Thus, a therapeutically effective amount can be determined by one of skill in the art of employing such practice in using no more than routine experimentation. For detailed information on HSC transplantations,

"Hemopoietic Stem Cell Transplantation, Its Foundation and Clinical Practice" (Modern Medicine, Special Issue, 53, 2, 1998) can be consulted and the descriptions given there are expressly incorporated herein by reference in their entirety.

[0101] In preparing the dosage of HSC to be administered, a variety of pharmaceutically acceptable carriers can be utilized. The carrier, diluent or vehicle may contain a buffering agent to obtain a physiologically acceptable H, such as phosphate- buffered saline, and/or other substances which are physiologically acceptable and/or are safe for use. In general, the material for intravenous injection in humans should conform to regulations established by the Food and Drug Administration, which are available to those in the field. Pharmaceutical ly-acceptable carriers may be combined, for example, in a 1 : 1 (v/v) ratio with the contacted HSC composition. The carrier may be for example, Ml 99 or RPMI 1640 medium. Furthermore, in preparing the dosage form, various infusions in common use today can also be employed. In one embodiment, the dose amount conventionally used in the transplantation of HSC can be employed. The dosage may be, for example, about 0.01-10 x 10 HSC/kg of weight of the patient, or more, or less where appropriate.

[0102] As noted above, the present invention provides pharmaceutical formulations comprising the compounds (including the pharmaceutically acceptable salts thereof), in pharmaceutically acceptable carriers for intravenous, oral, rectal, topical, buccal, parenteral, intramuscular, intradermal, and transdermal administration.

[0103] For in vivo embodiments in which a compound of the invention is delivered to a subject, the therapeutically effective dosage of any specific compound will vary somewhat from compound to compound, and subject to subject, and will depend upon the condition of the subject and the route of delivery. In general, a dosage from about 0.05 or 0.1 to about 20 or 50 mg/kg subject body weight may be utilized to carry out the present invention, e.g., about 0.05, 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 mg/kg or more or any range therein. For example, a dosage from about 0.1 mg/kg to about 50 mg/kg may be employed for oral administration; or a dosage of about 0.05 mg/kg to 20 mg/kg may be employed for intravenous injection. The compound may be delivered one or more times per day, e.g., 1, 2, 3, or 4 times a day. The compound may be delivered for a time sufficient to enhance the homing and/or engraftment of the HSC, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 18, 24, 48, or 72 hours or more.

[0104] The present invention is primarily concerned with the treatment of human subjects, but the invention may also be carried out on animal subjects, particularly mammalian subjects such as mice, rats, dogs, cats, livestock, and horses for veterinary purposes, and for drug screening and drug development purposes. The subjects may be male or female and may be of any suitable age, including infant, juvenile, adolescent, and adult subjects. In certain embodiments, the subject is one in need of the methods of the invention, e.g., in need of treatment or in need of transplantation.

IV. Compositions and Kits

[0105] One aspect of the invention relates to compositions comprising HSC that have been contacted with the compounds of the invention and optionally fucosylated. The HSC may be present in a physiologically or pharmaceutically acceptable carrier, e.g., saline or other formulations isotonic with the blood of the intended recipient. The compositions may contain other excipients suitable for delivery to a subject, such as antioxidants, buffers, and bacteriostats.

[0106] For in vivo embodiments of the invention, the compounds described above may be formulated for administration in a pharmaceutical carrier in accordance with known techniques. See, e.g., Remington, The Science And Practice of Pharmacy (21st Ed. 2006). In the manufacture of a pharmaceutical formulation according to the invention, the compound (including the physiologically acceptable salts thereof) is typically admixed with, inter alia, an acceptable carrier. The carrier must, of course, be acceptable in the sense of being compatible with any other ingredients in the formulation and must not be deleterious to the patient. The carrier may be a solid or a liquid, or both, and may be formulated with the compound as a unit-dose formulation, for example, a tablet, which may contain from 0.01 or 0.5% to 95% or 99% by weight of the compound. One or more compounds may be incorporated in the formulations of the invention, which may be prepared by any of the well known techniques of pharmacy consisting essentially of admixing the components, optionally including one or more accessory ingredients.

[0107] The formulations of the invention include those suitable for parenteral (e.g., intravenous, subcutaneous, intramuscular, or intradermal), oral, rectal, topical, buccal (e.g., sub-lingual), vaginal, topical (i.e., both skin and mucosal surfaces, including airway surfaces) and transdermal administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and on the nature of the particular compound which is being used.

[0108] Formulations of the present invention suitable for parenteral administration comprise sterile aqueous and non-aqueous injection solutions of the compound, which preparations are preferably isotonic with the blood of the intended recipient. These preparations may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient. Aqueous and non-aqueous sterile suspensions may include suspending agents and thickening agents. The formulations may be presented in unit\dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described. For example, in one aspect of the present invention, there is provided an injectable, stable, sterile composition comprising a compound of Formula I, or a salt thereof, in a unit dosage form in a sealed container. The compound or salt is provided in the form of a lyophilizate which is capable of being reconstituted with a suitable pharmaceutically acceptable carrier to form a liquid composition suitable for injection thereof into a subject. The unit dosage form typically comprises from about 10 μg to about 10 grams of the compound or salt. When the compound or salt is substantially water-insoluble, a sufficient amount of emulsifying agent which is physiologically acceptable may be employed in sufficient quantity to emulsify the compound or salt in an aqueous carrier. One such useful emulsifying agent is phosphatidyl choline.

[0109] Formulations suitable for oral administration may be presented in discrete units, such as capsules, cachets, lozenges, or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion. Such formulations may be prepared by any suitable method of pharmacy which includes the step of bringing into association the compound and a suitable carrier (which may contain one or more accessory ingredients as noted above). In general, the formulations of the invention are prepared by uniformly and intimately admixing the compound with a liquid or finely divided solid carrier, or both, and then, if necessary, shaping the resulting mixture. For example, a tablet may be prepared by compressing or molding a powder or granules containing the compound, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing, in a suitable machine, the compound in a free-flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, and/or surface active/dispersing agent(s). Molded tablets may be made by molding, in a suitable machine, the powdered compound moistened with an inert liquid binder.

[0110] Formulations suitable for buccal (sub-lingual) administration include lozenges comprising the compound in a flavored base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia.

[0111] Formulations suitable for rectal administration are preferably presented as unit dose suppositories. These may be prepared by admixing the compound with one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.

[0112] Formulations suitable for topical application to the skin preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil. Carriers which may be used include petroleum jelly, lanoline, polyethylene glycols, alcohols, transdermal enhancers, and combinations of two or more thereof.

[0113] Formulations suitable for transdermal administration may be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Formulations suitable for transdermal administration may also be delivered by iontophoresis {see, for example, Pharm. Res. 3(6):318 (1986)) and typically take the form of an optionally buffered aqueous solution of the compound. Suitable formulations comprise citrate or bis\tris buffer (pH 6) or ethanol/water and contain from 0.1 to 0.2M active ingredient.

[0114] Further, the present invention provides liposomal formulations of the compounds disclosed herein and salts thereof. The technology for forming liposomal suspensions is well known in the art. When the compound or salt thereof is an aqueous- soluble salt, using conventional liposome technology, the same may be incorporated into lipid vesicles. In such an instance, due to the water solubility of the compound or salt, the compound or salt will be substantially entrained within the hydrophilic center or core of the liposomes. The lipid layer employed may be of any conventional composition and may either contain cholesterol or may be cholesterol-free. When the compound or salt of interest is water-insoluble, again employing conventional liposome formation technology, the salt may be substantially entrained within the hydrophobic lipid bilayer which forms the structure of the liposome. In either instance, the liposomes which are produced may be reduced in size, as through the use of standard sonication and homogenization techniques. Liposomal formulations containing the compounds disclosed herein or salts thereof, may be lyophilized to produce a lyophilizate which may be reconstituted with a pharmaceutically acceptable carrier, such as water, to regenerate a liposomal suspension.

[0115] Other pharmaceutical compositions may be prepared from the water-insoluble compounds disclosed herein, or salts thereof, such as aqueous base emulsions. In such an instance, the composition will contain a sufficient amount of pharmaceutically acceptable emulsifying agent to emulsify the desired amount of the compound or salt thereof. Particularly useful emulsifying agents include phosphatidyl cholines, and lecithin.

[0116] In addition to the compounds, the pharmaceutical compositions may contain other additives, such as pH-adjusting additives. In particular, useful pH-adjusting agents include acids, such as hydrochloric acid, bases or buffers, such as sodium lactate, sodium acetate, sodium phosphate, sodium citrate, sodium borate, or sodium gluconate. Further, the compositions may contain microbial preservatives. Useful microbial preservatives include methylparaben, propylparaben, and benzyl alcohol. The microbial preservative is typically employed when the formulation is placed in a vial designed for multidose use. Of course, as indicated, the pharmaceutical compositions of the present invention may be lyophilized using techniques well known in the art.

[0117] Another aspect of the invention relates to kits comprising the HSC and/or compounds of the invention. The kits may further contain other reagents for enhancing HSC homing and/or engraftment, such as an antibody which binds a specific type of stem cell, reagents for fucosylation, a vessel for incubation of stem cells, means for administering stem cells to a patient, a pharmaceutically acceptable carrier, a physiologically acceptable carrier, instructions for use, a container, a vessel for administration, an assay substrate, or any combination thereof.

[0118] The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.

Claims

That which is claimed is:

1. A method for increasing homing and/or engraftment of hematopoietic stem cells (HSC) to bone marrow comprising contacting HSC with an effective amount of a compound of Formula I:

wherein:

X is NR³ or O;

n is 1 or 2;

A is a bicyclic or tricyclic carbocycle of 5 to 20 atoms wherein each bridge bicycle has at least one atom;

R¹ is

wherein:

p and q are independently 0 or 1 ;

Y is CH₂, CHF, CF₂, O, or S(0)_m;

W and Z are independently CH₂, CHF, or CF₂;

when X is NR³ then R² is R⁴-S0₂-; R⁵-S0₂-NH-C(0)-; R⁶R⁷N-S0₂-; or a heterocyclic group unsubstituted or optionally substituted with halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, haloalkyloxy, cycloalkyloxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocycloalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, cycloalkyl-S(0)_m, cycloalkylalkyl-S(0)_m, aryl-S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano; or when X is O then R² is a heterocyclic group unsubstituted or optionally substituted with halo, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, heterocycloalkyl, hydroxyl, alkoxy, haloalkyloxy, cycloalkyloxy, cycloalkylalkyloxy, aryloxy, arylalkyloxy, heterocyclooxy, heterocycloalkyloxy, mercapto, alkyl-S(0)_m, haloalkyl-S(0)_m, cycloalkyl-S(0)_m, cycloalkylalkyl-S(0)_m, aryl- S(0)_m, arylalkyl-S(0)_m, heterocyclo-S(0)_m, heterocycloalkyl-S(0)_m, amino, alkylamino, haloalkylamino, cycloalkylamino, cycloalkylalkylamino, arylamino, arylalkylamino, heterocycloamino, heterocycloalkylamino, disubstituted-amino, acylamino, acyloxy, ester, amide, sulfonamide, urea, alkoxyacylamino, aminoacyloxy, nitro or cyano;

R⁸ is H or cyano;

m is 0, 1 or 2;

or a pharmaceutically acceptable salt or prodrug thereof.

2. The method of claim 1, wherein A is selected from the group consisting of adamantyl, bicyclo[2.1.1]hexane, bicyclo[3.1.1]heptane, bicyclo[3.2.1]octane, bicyclo[2.2.2]octane, and bicyclo[3.3.1]nonane, each of which may optionally include one or more double bonds.

3. The method of claim 1, wherein A is selected from the group consisting of:

4. The method of claim 1, wherein X is NR³ and R² is R⁴-S0₂-, R⁵-S0₂-NH-C(0)-, R⁶R⁷N-S0₂- or a heterocyclic group.

5. The method of claim 1, wherein X is O and R² is a heterocyclic group.

6. The method of claim 1, wherein the compound is selected from the consisting of:

and pharmaceutically acceptable salts and prodrugs thereof. 7. The method of claim 1, wherein the compound is:

a pharmaceutically acceptable salt or prodrug thereof. The method of claim 1, wherein the compound is:

pharmaceutically acceptable salt or prodrug thereof.

The method of claim 1, wherein the compound is:

or a pharmaceutically acceptable salt or prodrug thereof.

10. The method of claim 1, wherein:

Y is selected from the group consisting of CHF, CF₂, O, and S(0)_m; or

q is 1 and W is selected from the group consisting of CHF and CF₂; or

p is 1 and Z is selected from the group consisting of CHF and CF₂.

11. The method of claim 1 , wherein:

Y is selected from the group consisting of CHF, CF₂, O, and S(0)_m;

q is 1 and W is CH₂; and

p is 0.

12. The method of claim 1, wherein:

Y is selected from the group consisting of CHF, CF₂, O, and S(0)_m; or

q is 0; and

p is 1 and Z is CH₂.

13. The method of claim 1, wherein:

Y is CH₂;

q is 1 and W is selected from the group consisting of CHF and CF₂; and

p is 0.

14. The method of claim 1, wherein:

Y is CH₂;

q is 0; and

p is 1 and Z is selected from the group consisting of CHF and CF₂.

15. The method of claim 1, wherein the compound is selected from the group consisting of:

-40-

-41 -

-42-

-43 -

-44-

-45-

-46-

-47-

and pharmaceutically acceptable salts and prodrugs thereof.

16. The method of claim 1, wherein the compound is selected from the group consisting of:

and pharmaceutically acceptable salts and prodrugs thereof.

17. The method of claim 1, wherein said HSC are contacted in vitro or ex vivo.

18. The method of claim 1, wherein said HSC are isolated from a subject prior to contacting.

19. The method of claim 1, wherein said HSC are contacted in vivo in a subject.

20. The method of claim 19, wherein said compound is delivered to the subject before, during, and/or after delivery of HSC to the subject.

21. The method of claim 1, wherein said HSC are contacted both in vitro or ex vivo and in vivo in a subject.

22. The method of claim 1, wherein said HSC are from umbilical cord blood.

23. The method of claim 1, wherein said HSC are from peripheral blood.

24. The method of claim 1, wherein said HSC are from bone marrow.

25. The method of claim 1, wherein said HSC are human cells.

26. The method of claim 1, further comprising fucosylating the HSC.

27. A composition comprising HSC produced by the method of claim 1.

28. The composition of claim 27, further comprising a physiologically acceptable carrier.

29. A method for treating a condition in a subject for which transplantation of HSC is required or advantageous, comprising:

providing the composition of claim 27, and

delivering to the subject a therapeutically effective quantity of the composition.

30. The method of claim 29, wherein the condition is selected from the group consisting of chronic myelocytic (myelogenous) leukemia, juvenile chronic myelogenous leukemia, chronic lymphocytic leukemia, acute myelocytic leukemia, acute lymphocytic leukemia, malignant lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, multiple myeloma, aplastic anemia gravis, sickle cell anemia, myelodysplastic syndrome, autoimmune disease, Gunther's disease, Hunter syndrome, Hurler syndrome, neuroblastoma, Wiskott-Aldrich syndrome, X-linked lympho-proliferative syndrome, and solid tissue tumors.

31. A method for performing a bone marrow transplantation in a subject, comprising: providing the composition of claim 27, and

32. The method of claim 29 or 31, further comprising delivering to said subject a compound of Formula I:

wherein:

X is NR³ or O;

n is 1 or 2; A is a bicyclic or tricyclic carbocycle of 5 to 20 atoms wherein each bridge of the bicycle has at least one atom;

R¹ is

wherein:

p and q are independently 0 or 1;

Y is CH₂, CHF, CF₂, O, or S(0)_m;

W and Z are independently CH₂, CHF, or CF₂;

R³ is selected from the group consisting of H, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl; R is selected from the group consisting of haloalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heterocyclo, and heterocycloalkyl;

R⁸ is H or cyano;

m is 0, 1 or 2;

or a pharmaceutically acceptable salt or prodrug thereof.