Claims for Patent: 7,887,833

Summary for Patent: 7,887,833

Title:	Amphiphilic compounds and vesicles liposomes for organ-specified drug targeting
Abstract:	An amphiphilic compound is provided capable of forming vesicles or liposomes, said amphiphilic compound having at least one headgroup containing a selectively cleavable group or moiety such as a residue of a choline or phenylalanine derivative, and at least one hydrogen-bonding group located either within said headgroup and/or in close proximity thereto. The cleavable group or moiety is cleaved under selective conditions including change of chemical, physical or biological environment and is preferably cleaved enzymatically in a biological environment such as the brain or the blood. Vesicles or liposomes made from said amphiphilic compounds are suitable for delivery of a therapeutic substance or a diagnostic agent specifically to a target organ or tissue, or for delivery of a nucleic acid for gene therapy.
Inventor(s):	Heldman; Eliahu (Rehovot, IL), Linder; Charles (Rehovot, IL), Grinberg; Sarina (Meitar, IL), Kolot; Victoria (Beer-Sheva, IL), Shaubi; Eleonora (Beer-Sheva, IL)
Assignee:	Ben-Gurion University of the Negev Research and Development Authority (Beer-Sheva, IL)
Application Number:	10/497,705
Patent Claims:	1. An amphiphilic compound capable of forming vesicles or liposomes, of the formula Ia: R.sub.8--R.sub.7-A.sub.4-R.sub.6-A.sub.3-R.sub.5--OC-Q.sub.2-R.sub.10-Q.s- ub.1-CO--R.sub.1-A.sub.1-R.sub.2-A.sub.2-R.sub.3--R.sub.4 (Ia) wherein: R.sub.1 and R.sub.5, the same or different, each is --(CH.sub.2).sub.n; A.sub.1 is selected from the group consisting of --(CH.sub.2).sub.m+2--, --CH.dbd.CH--(CH.sub.2).sub.m--, --CH.dbd.CH--CH(Y.sub.1)--, --CH.sub.2--CH.sub.2--CH(Y.sub.1)--, (--CH.sub.2--CH(Y.sub.1)--(CH.sub.2).sub.m--, --CH(Y.sub.1)--CH.sub.2--(CH.sub.2).sub.m--, --CH(Y.sub.1)--CH(Y.sub.2)--(CH.sub.2).sub.m--, wherein Y.sub.1 and Y.sub.2 each is halogen, --OH, --O--CO--(CH.sub.2).sub.m--Y.sub.3, --NH--CO--Y.sub.3, --SH, --SR.sub.11, --NH.sub.2, and --N(R.sub.11)(R.sub.12), or Y.sub.1 and Y.sub.2 together with the carbon atoms to which they are attached form a 2,3-oxiranylene group; and Y.sub.3 is halogen, --OH, --SH, --NH.sub.2, or --N(R.sub.11)(R.sub.12); R.sub.2 and R.sub.6, the same or different, each is C.sub.1-C.sub.4 alkylene, unsubstituted or substituted by halogen, amino or hydroxy; A.sub.2 is --CH(R.sub.13)--, --CH.sub.2--CH(R.sub.13)--, --CH(R.sub.13)--CH.sub.2--, --CH(OH)--CH(R.sub.13) --, --CH(R.sub.13)--CH(OH)--, --CH(OH)--CH.sub.2--CH(OH)--CH(R.sub.13)--, --CH(OH)--CH.sub.2--CH(R.sub.13)--CH(OH)--, -G1-(C.sub.6-C.sub.14 arylene)-(CH.sub.2).sub.qR.sub.14, --N(CH.sub.3).sub.2R.sub.14, or --SR.sub.14; R.sub.3 and R.sub.7, the same or different, each is --(CH.sub.2).sub.o--; R.sub.4 is H or CH.sub.3, and wherein the total sum of carbon atoms in the R.sub.1-A.sub.1-R.sub.2-A.sub.2-R.sub.3--R.sub.4 chain is at most 23; Q.sub.1 is --NH--, --O--, --S--, or --O--PO(OH)--O--; Q.sub.2 is --NH--, --O--, --S--, or --O--PO(OH)--O--; R.sub.10 is --(CH.sub.2).sub.p--; --CH.sub.2(CH.sub.3)--(CH.sub.2).sub.p--; --CH(CH.sub.3)--(CH.sub.2).sub.p--CH(CH.sub.3)--; --(CH.sub.2--CH.sub.2--O--).sub.p--CH.sub.2--CH.sub.2--; --(CH.sub.2--CH.sub.2--S--).sub.p--CH.sub.2--CH.sub.2--; --(CH.sub.2--CH.sub.2--NH--).sub.p--CH.sub.2--CH.sub.2--; --C.sub.6-C.sub.14 arylene-; --(C.sub.6-C.sub.14 arylene)-R--(C.sub.6-C.sub.14 arylene), wherein R is C.sub.1-C.sub.4 alkylene, --CCH.sub.3).sub.2--, --O--, --S--, --NH-- or --SO.sub.2--; A.sub.3 is as defined for A.sub.1, or is --(CH.sub.2).sub.m, phenyl or --CH.sub.2-phenyl, wherein the phenyl ring is unsubstituted or substituted by C.sub.1-C.sub.4 alkyl, halogen or both; A.sub.4 is as defined for A.sub.2, or is --(CH.sub.2).sub.m--; R.sub.8 is as defined for R.sub.4; R.sub.13 is -G1-(CH.sub.2).sub.mR.sub.14 or -G1-CO(CH.sub.2).sub.mR.sub.14; G1 is --O--, --S--, --NR''--, --CH2NR''--, --CH2S--, --CH2O--, --NH--CO--, --O--CO--NH--, --NH--CO--NH--, --C.dbd.NO--, or --C(NH2)=NO--, wherein R'' is H or C.sub.1-C.sub.18 alkyl; R.sub.14 is either a headgroup containing a selectively cleavable group or moiety, or is as defined for R.sub.15 or for R.sub.15 substituted by a selectively cleavable group or moiety; R.sub.11 and R.sub.12, the same or different, each is C.sub.1-C.sub.18 alkyl unsubstituted or substituted by halogen; phenyl or --CH.sub.2-phenyl, wherein the phenyl ring is unsubstituted or substituted by C.sub.1-C.sub.4-alkyl, halogen, or both, and wherein one of R.sub.11 and R.sub.12 may be H; R.sub.15 is --NH.sub.2; --NR.sub.11R.sub.12; --N.sup.+R.sub.11R.sub.12R.sub.16 wherein R.sub.16 is as defined for R.sub.11 and R.sub.12; --O--CO--(C.sub.2-C.sub.6 alkenyl); --O--CO--(CH.sub.2).sub.t--NR.sub.11R.sub.12; --O--CO--(CH.sub.2).sub.t--N.sup.+R.sub.11R.sub.12R.sub.16; --O--CO--(CH.sub.2).sub.t--COOH; --O--CO--(CH.sub.2).sub.t--SO.sub.3H; --O--CO--(CH.sub.2).sub.t--O--PO(OH).sub.2; --NH--(CH.sub.2).sub.r--COOH; --NH--(CH.sub.2).sub.r--SO.sub.3H; --NH--(CH.sub.2).sub.r--O--PO(OH).sub.2; --NH--PO(OH).sub.2; --N.sup.+(CH.sub.3).sub.2--R.sub.17; --O--PO(OH)--O--(CH.sub.2).sub.2--N.sup.+R.sub.11R.sub.12R.sub.16; --O--PO(OH)--O--(CH.sub.2).sub.2--NH.sub.3.sup.+; --O--PO(OH)--NH--PO(OH)--O--; --O--PO(OH)--O--CH.sub.2--CH(NH.sub.3.sup.+)--COO.sup.-; --CH.sub.2--CH.dbd.CH.sub.2; --CO--CH.dbd.CH.sub.2; --CO--C(CH.sub.3).dbd.CH.sub.2; --(CH.sub.2).sub.r--COOH; --(CH.sub.2).sub.r--O--SO.sub.2H; --(CH.sub.2).sub.r--O--PO(OH).sub.2; --SR.sub.18; -G1-(C.sub.6-C.sub.14 arylene)-NR.sub.11R.sub.12; -G1-(C.sub.6-C.sub.14 arylene)-N.sup.+R.sub.11R.sub.12R.sub.16; -G1-(C.sub.6-C.sub.14 arylene)-COOH; -G1-(C.sub.6-C.sub.14 arylene)-SO.sub.3H; -G1-(C.sub.6-C.sub.14 arylene)-O--PO(OH).sub.2; -G1-(C.sub.6-C.sub.14 arylene)-(CH.sub.2).sub.t--NR.sub.11R.sub.12; -G1-(C.sub.6-C.sub.14 arylene)-(CH.sub.2).sub.n--N.sup.+R.sub.11R.sub.12R.sub.16; -G1-(C.sub.6-C.sub.14 arylene)-(CH.sub.2).sub.t--COOH; or -G1-(C.sub.6-C.sub.14 arylene)-(CH.sub.2).sub.t--SO.sub.3H; R.sub.17 is --CH.sub.2--CH.dbd.CH.sub.2, --CO--CH.dbd.CH.sub.2, --CO--C(CH.sub.3).dbd.CH.sub.2, --(CH.sub.2).sub.q--N.sup.+R.sub.11R.sub.12R.sub.16, --(CH.sub.2).sub.q--NH--(CH.sub.2).sub.q--SO.sub.3H, --(CH.sub.2).sub.q--NH--(CH.sub.2).sub.q--COOH, --(CH.sub.2).sub.q--NH--(CH.sub.2).sub.q--O--PO(OH).sub.2, --PO(OH).sub.2, or --O--PO(OH)--O--(CH.sub.2).sub.2--N.sup.+R.sub.11R.sub.12R.sub.16; R.sub.18 is hydrogen, C.sub.1-C.sub.18 alkyl, C.sub.2-C.sub.6 alkenyl with a terminal double bond, --CO--CH.dbd.CH.sub.2, or --CO--C(CH.sub.3).dbd.CH--NR.sub.11R.sub.12; n is an integer from 5 to 10; m is an integer from 0 to 4; o is an integer from 0 to 10; p is an integer from 1 to 16; q is an integer from 0 to 3; r is an integer from 1 to 6; and t is an integer from 1 to 14, and salts thereof; wherein said amphiphilic compound of formula Ia has at least one radical R.sub.14 which is a headgroup containing a selectively cleavable group or moiety and at least one hydrogen-bonding group located within said at least one headgroup, in close proximity thereto, or within and in close proximity to said at least one headgroup; and said selectively cleavable group or moiety is a group or moiety that is cleaved under selective conditions selected from the group consisting of change of chemical conditions, physical conditions, biological environment and combinations thereof. 2. An amphiphilic compound according to claim 1, wherein said selective conditions are selected from the group consisting of change of pH, change of temperature, oxidative conditions, reducing conditions, enzymatic conditions, and combinations thereof. 3. An amphiphilic compound according to claim 1, wherein said cleavable group is cleaved enzymatically in a biological environment. 4. An amphiphilic compound according to claim 3, wherein said biological environment is the brain or the blood. 5. An amphiphilic compound according to claim 1, wherein said selectively cleavable group or moiety is a residue of a derivative of a compound selected from the group consisting of choline, an aromatic amino acid, a dicarboxylic amino acid, a saccharide and a peptide that is specifically cleaved by an enzyme at a desired site. 6. An amphiphilic compound according to claim 5, wherein said derivative of choline is acetylcholine or acetylthiocholine; said aromatic amino acid is selected from the group consisting of phenylalanine, tyrosine, tryptophan and a derivative thereof selected from the group consisting of p-aminophenylalanine and levodopa; said dicarboxylic amino acid is glutamic acid or aspartic acid; said saccharide is glucose, galactose or mannose; and said peptide is enkephaline or N-acetyl-ala,ala. 7. An amphiphilic compound according to claim 1, wherein said hydrogen-bonding group is selected from the group consisting of --OH, --SH, --NH--, --N.sup.+H.sub.2--, --NH.sub.2, --N.sup.+H.sub.3, --NH--CO--, --O--CO--NH--, --NH--CO--NH--, --C.dbd.NOH, --C(NH.sub.2).dbd.NOH, --C(NH.sub.2).dbd.NO-- and --CO--NH.sub.2. 8. An amphiphilic compound according to claim 1, capable of forming monolayer vesicles, of the formula Ia: R.sub.8--R.sub.7-A.sub.4-R.sub.6-A.sub.3-R.sub.5--OC-Q.sub.2-R.sub.10-Q.s- ub.1-CO--R.sub.1-A.sub.1-R.sub.2-A.sub.2-R.sub.3--R.sub.4 (Ia) wherein: R.sub.1 and R.sub.5 are each --(CH.sub.2).sub.7; A.sub.1 is --CH.dbd.CH--(CH.sub.2).sub.m--, wherein m is 0; R.sub.2 and R.sub.6 each is --CH.sub.2--; A.sub.2 is --CH(OH)--CH(R.sub.13)--; R.sub.3 and R.sub.7 each is --(CH.sub.2).sub.4--; R.sub.4 is CH.sub.3, and wherein the total sum of carbon atoms in the R.sub.1-A.sub.1-R.sub.2-A.sub.2-R.sub.3--R.sub.4 chain is 17; Q.sub.1 is --NH--; Q.sub.2 is --NH--; R.sub.10 is --(CH.sub.2).sub.2--; A.sub.3 is as defined for A.sub.1; A.sub.4 is as defined for A.sub.2; R.sub.8 is as defined for R.sub.4; R.sub.13 is --O--COCH.sub.2--R.sub.14; and R.sub.14 is a headgroup containing the selectively cleavable group or moiety --N.sup.+(CH.sub.3).sub.2--CH.sub.2--CH.sub.2--O--CO--CH.sub.3; and salts thereof. 9. An amphiphilic compound according to claim 1, selected from the group consisting of: R.sub.19--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.19 (Ib), R.sub.19--CO--O--(CH.sub.2).sub.2--O--CO--R.sub.19 (Ic), R'.sub.19--CO--NH--(CH.sub.2).sub.2--NH--CO--R'.sub.19 (Id) R'.sub.19--CO--NH--(CH.sub.2).sub.2--NH--CO--R'.sub.19 (Ie), R.sub.24--CO--NH--(CH.sub.2).sub.2--NH--CO--R'.sub.19 (If), and R.sub.27--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.26 (Ig), wherein R.sub.19 is --(CH.sub.2).sub.7--CH.dbd.CH--CH.sub.2--CH(OH)--CH(R.sub.20)--(CH.sub.2)- .sub.4--CH.sub.3, R'.sub.19 is --(CH.sub.2).sub.7--CH.dbd.CH--CH.sub.2--CH(OH)--CH(R.sub.21)--(CH.sub.2)- .sub.4--CH.sub.3, R''.sub.19 is --(CH.sub.2).sub.7--CH.dbd.CH--CH.sub.2--CH(OH)--CH(R.sub.23)--(CH.sub.2)- .sub.4--CH.sub.3, R.sub.20 is --OCOCH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH.sub.2)--COOH, R.sub.21 is --NHCO--CH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH.sub.2)--COOH, R.sub.23 is --NH--CH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH.sub.2)--COOH, R.sub.24 is --(CH.sub.2).sub.7--CH.dbd.CH--CH.sub.2--CH(OH)--CH(R.sub.25)--(CH.sub- .2).sub.4--CH.sub.3, R.sub.25 is --NH--CO--CH.sub.2--COOH, and R.sub.26 and R.sub.27 is each --(CH.sub.2).sub.12--CH(OH)--CH.sub.2--R.sub.23. 10. An amphiphilic compound according to claim 1, selected from the group consisting of: R.sub.28--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.28 (Ih), R.sub.29--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.29 (Ii), R.sub.31--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.31 (Ik), R.sub.33--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.31 (Il), R.sub.35--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.35 (Im), R.sub.37--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.37 (In), R.sub.39--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.39 (Io), and R.sub.41--CO--NH--(CH.sub.2).sub.2--NH--CO--R.sub.41 (Ip), wherein R.sub.28 is --(CH.sub.2).sub.12--NHCH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH.sub.2)CO- OH, R.sub.29 is --(CH.sub.2).sub.12--NHCO--CH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH.sub.- 2)COOH, R.sub.31 is --(CH.sub.2).sub.12--R.sub.32, R.sub.32 is --NHCH.sub.2CH.sub.2N.sup.+(CH.sub.3).sub.2--CH.sub.2--CH.sub.2--OCOCH.su- b.3, R.sub.33 is --(CH.sub.2).sub.12--R.sub.34, R.sub.34 is --NHCH.sub.2CH.sub.2N.sup.+(CH.sub.3).sub.3, R.sub.35 is --(CH.sub.2).sub.9--CH.dbd.CH--CH.sub.2--CH(R.sub.36)--(CH.sub.2).sub.5CH- .sub.3, R.sub.36 is --OCH.sub.2CH.sub.2NH--CO--CH.sub.2--CH.sub.2--CH(NH.sub.2)COOH, R.sub.37 is --(CH.sub.2).sub.7--CH.dbd.CH--CH.sub.2--CH(R.sub.38)--(CH.sub.2).sub.- 5CH.sub.3, R.sub.38 is --OCH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH.sub.2)COOH, R.sub.39 is --(CH.sub.2).sub.12--N(R.sub.40)CH.sub.2CH.sub.2NH-phenyl-CH.sub.2--CH(NH- .sub.2)COOH R.sub.40 is a C.sub.4-C.sub.16 alkyl, and R.sub.41 is --(CH.sub.2).sub.7--CH.dbd.CH--CH.sub.2--CH(OR.sub.40)--CH(R.sub.38)--(CH- .sub.2).sub.4--CH.sub.3. 11. Vesicles or liposomes made from at least one amphiphilic compound according to claim 1. 12. Vesicles or liposomes according to claim 11, presenting one or more of the following characteristics: (i) an ionic or polar headgroup attached to a hydrophobic chain, said headgroup containing a selectively cleavable group or moiety and a hydrogen-bonding group within the headgroup or in close proximity thereto; (ii) two ionic or polar headgroups on opposite ends of the hydrophobic chain moiety, at least one containing a selectively cleavable group or moiety and a hydrogen-bonding group within the headgroup or in close proximity thereto, and optionally containing additional ionic and polar groups in proximity to the said headgroups, that do not hinder vesicle formation but which are capable of polar or hydrogen-bonding interactions that stabilize the vesicles which are made therefrom; (iii) two headgroups on opposite ends of the molecule, at least one containing a selectively cleavable group or moiety and a hydrogen-bonding group within the headgroup or in close proximity thereto, and said headgroups may optionally contain additional polar groups within the center or in relative proximity to the midsection or center of the hydrophobic chain, capable of polar or hydrogen-bonding interactions that stabilize the vesicles which are made therefrom; and (iv) in addition to (i) to (iii), additional aliphatic chain(s) of at least 5 methylene groups attached to the aforementioned hydrophobic chain moiety or to the headgroups to increase the hydrophobic interactions, and thus, contribute to the stability of vesicles which are made therefrom. 13. Vesicles or liposomes according to claim 11, being monolayered nanovesicles with asymmetric headgroups, made from double headed amphiphilic derivatives with two different polar or ionic headgroups on opposite sides of the aliphatic chains (bolaamphiphiles), wherein at least one of the said headgroups (the bulkier one) contains a group or moiety located on the outer vesicle surface, that can be selectively cleaved, and additional polar or ionic headgroups on the aliphatic chain(s) that give the vesicle additional stability, but nevertheless allow the vesicle to disrupt after removal of the ionic or polar headgroups. 14. Vesicles or liposomes according to claim 11, comprising a therapeutic substance encapsulated in, or loaded on, the vesicle or liposome. 15. Vesicles or liposomes according to claim 14, wherein said therapeutic substance is selected from the group consisting of levodopa (L-DOPA), a combination of carbidopa and levodopa, apomorphine, dopamine, and glial derived neurotrophic factor. 16. Vesicles or liposomes according to claim 14, wherein said therapeutic substance is a chemotherapeutic drug for chemotherapy of a brain tumor. 17. Vesicles or liposomes according to claim 14, for delivery of a therapeutic substance to the brain wherein said vesicles or liposomes further comprising amphiphilic compounds with headgroups containing targeting groups to the brain, groups for transport through the blood brain barrier, groups to stabilize the vesicles against clearance by the body, or combinations thereof. 18. Vesicles or liposomes according to claim 15, wherein said targeting groups to the brain are selected from the group consisting of residues of nicotine, cytosine, lobeline, L-glutamic acid, MK801, morphine, enkephaline, benzodiazepines, dopamine antagonists, tricyclic antidepressants, muscarinic agonists, muscarinic antagonists and cannabinoids. 19. Vesicles or liposomes according to claim 17, wherein said group for transport through the blood brain barrier is selected from the group consisting of glucose, mannose, and ascorbic acid, an antibody, choline, or an amino acid selected from the group consisting of glutamic acid, tryptophan and levodopa. 20. Vesicles or liposomes according to claim 17, wherein said groups to stabilize the vesicles against clearance by the body are polyethyleneglycol (PEG) residues. 21. Vesicles or liposomes according to claim 14, wherein said therapeutic substance has a short lifetime at the delivery site and exerts its activity in another part of the body. 22. Vesicles or liposomes according to claim 21, wherein said therapeutic substance is selected from the group consisting of insulin, copaxone, herceptin and a mixture of immunoglobulins. 23. The amphiphilic compounds herein designated as Derivatives 1, 2, 3, 4, 5, 8, 9 and 12 having the formulas ##STR00054## ##STR00055## 24. The amphiphilic compounds herein designated as Derivatives 6, 10 and 11, as described at pages 68, 45 and 72 of the specification, respectively. 25. Vesicles or liposomes according to claim 14, wherein said therapeutic substance is selected from the group consisting of an antibody against components of Alzheimer plaques, an anti-inflammatory agent, a growth factor, and a muscarinic agonist.

Details for Patent 7,887,833

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	09/25/1998		2039-02-26
Genentech, Inc.	HERCEPTIN	trastuzumab	For Injection	103792	02/10/2017		2039-02-26