Ortho ester lipids

• US 6,200,599 B1
• Filed: 10/07/1999
• Issued: 03/13/2001
• Est. Priority Date: 10/07/1999
• Status: Expired due to Fees

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Zhu, Ji, Nantz, Michael H.
• Primary Examiner(s): Schwartzman, Robert A.
• Assistant Examiner(s): Ousley, Andrea
• Application Number:
  US09/415,016
• Time in Patent Office: 523 Days
• Field of Search: 435/440, 435/458, 424/450
• US Class Current: 424/450
• CPC Class Codes: A61K 9/1272 with substantial amounts of non-phosphatidyl, i.e. non-acylglycerophosphate, surfactants as bilayer-forming substances, e.g. cationic lipids with cholesterol as the only non-phosphatidyl surfactant A61K9/127; lipids as modifying agent A61K47/543
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 23/02 Local anaesthetics
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/18 Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
• CPC Class Codes: A61P 25/20 Hypnotics; Sedatives
• CPC Class Codes: A61P 25/22 Anxiolytics
• CPC Class Codes: A61P 25/24 Antidepressants
• CPC Class Codes: A61P 25/36 Opioid-abuse
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 29/02 without antiinflammatory effect
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 3/14 for calcium homeostasis vitamin D A61P3/02; parathyroid hormones A61P5/18; calcitonin A61P5/22; osteoporosis A61P19/10; bone metastasis A61P35/04
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 : Antimycotics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/08 : Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 5/18 : of the parathyroid hormones
• CPC Class Codes: A61P 7/02 : Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
• CPC Class Codes: A61P 9/04 : Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: A61P 9/12 : Antihypertensives
• CPC Class Codes: A61P 9/14 : Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
• CPC Class Codes: C07D 493/08 : Bridged systems

Abstract

The present invention provides ortho ester lipids and their derivatives that, upon certain pH conditions, undergo hydrolysis with concomitant or subsequent headgroup cleavage. These ortho ester lipids can advantageously be formulated into liposomes. The liposome formulations are useful in nucleic acid transfection and entrapment/delivery of conventional small molecules and therapeutic agents. Moreover, the liposomes comprising compounds of the present invention are useful as drug delivery vehicles.

Claims

1. A compound having the formula

wherein:

R¹ is a member selected from the group consisting of optionally substituted (C₇-C₁₇)alkyl, optionally substituted (C₇-C₁₇)alkenyl and optionally substituted (C₇-C₁₇)alkynyl;

R² is a member selected from the group consisting of (C₁-C₁₈)alkoxy and (C₁-C₁₈)alkylthio;

R³ is hydrogen;

alternatively,

R² and R³ and the carbons to which they are bound join to form a 5,6-membered;

a 6,6-membered;

a 6,7-membered;

or a 7,7-membered bicyclic ortho ester or ortho thioester ring;

A and A¹ are members independently selected from the group consisting of oxygen and sulfur;

x and y are independently selected from the integers 0, 1 and 2;

Z is a member selected from the group consisting of optionally substituted alkylene, optionally substituted alkyleneoxyalkylene and optionally substituted alkyleneaminoalkylene;

Q is a member selected from the group consisting of carboxyl, thiocarboxyl, dithiocarboxyl, carbonate, carbamate, phospho, phosphothio, phosphoro and thiophosphoro; and

R⁴ is a nitrogen containing headgroup wherein the nitrogen can be unsubstituted, mono-substituted, di-substituted, or a quaternary nitrogen salt and wherein the nitrogen substituent(s) are member(s) independently selected from the group consisting of optionally substituted (C₁-C₁₈)alkyl, optionally substituted (C₁-C₁₈)alkenyl, and optionally substituted (C₁-C₁₈)alkynyl and wherein R⁴ and Q are optionally linked with a (C₁-C₅)alkylene or (C₂-C₅)alkenyl group.

2. The compound of claim 1, wherein:

R¹ is a member selected from the group consisting of (C₇-C₁₇)alkyl and substituted (C₇-C₁₇)alkyl;

R² and R³ and the carbons to which they are bound join to form a 6,6-membered;

a 6,7-membered;

or a 7,7-membered bicyclic ortho ester ring;

A and A¹ are oxygen;

x and y are independently selected from the integers 0, 1 and 2;

Z is optionally substituted alkylene;

Q is a member selected from the group consisting of carboxyl, phospho, and phosphoro;

R⁴ is a member selected from the group consisting of optionally substituted amino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)alkoylamino(C₂-C₅)alkenyl, optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)dialkylamino(C₂-C₅)alkenyl, and

a quaternary nitrogen salt having the structure

3. The compound of claim 1, having the formula

wherein:

R¹ is a member selected from the group consisting of (C₇-C₁₇)alkyl and substituted (C₇-C₁₇)alkyl;

Z is optionally substituted alkylene;

Q is carboxyl;

R⁴ is a member selected from the group consisting of optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene and a quaternary nitrogen salt having the structure

4. The compound of claim 3, wherein:

R¹ is (C₇-C₁₇)alkyl optionally substituted with one or more members selected from the group consisting of lower alkyl, acyl, halogen, hydroxy, amino, alkoxy, alkylamino, acylamino, acyloxy, and mercapto;

Z is (C₁-C₃) alkylene optionally substituted with one or more members selected from the group consisting of lower alkyl, aryl, acyl, halogen, hydroxy, amino, alkoxy, alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, mercapto, both saturated and unsaturated cyclic hydrocarbons and heterocycles;

R⁴ is a member selected from the group consisting of optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene or optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene, wherein the substituent is one or more members selected from the group consisting of lower alkyl, aryl, acyl, halogen, hydroxy, amino, alkoxy, alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, mercapto, both saturated and unsaturated cyclic hydrocarbons and heterocycles; and

an ammonium salt having the structure;

5. The compound of claim 4, wherein:

R¹ is (C₇-C₁₇)alkyl optionally substituted with (C₇-C₁₈)acyloxy;

Z is (C₁-C₃) alkylene optionally substituted with (C₇-C₁₈)alkoxy; and

R⁴ is a member selected from the group consisting of optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene wherein the substituent is (C₁-C₁₈)acyloxy, and an ammonium salt wherein;

R⁶ is methyl.

6. The compound of claim 3, wherein:

R¹ is a (C₁₃-C₁₇)alkyl;

Z is (C₁-C₃)alkylene;

R⁴ is a member selected from the group consisting of (C₁₄-C₁₈)acyloxy(C₁-C₅)alkylamino(C₂-C₄)alkylene and an ammonium salt wherein;

R⁵ and R⁶ are both methyl; and

R⁷ is (C₁₄-C₁₈)acyloxy(C₁-C₅)alkylene.

7. The compound of claim 3, wherein:

R¹ is (C₁₄)alkyl;

Z is (C₂)alkylene; and

R⁴ is a member selected from the group consisting of (C₁₄)alkylamino(C₂)alkylene and an ammonium salt wherein;

R⁵ and R⁶ are both methyl; and

R⁷ is (C₁₄)alkyl.

8. The compound of claim 3, wherein:

R¹ is (C₁₄)alkyl;

Z is (C₂)alkylene; and

R⁴ is a member selected from the group consisting of N-methyl-N-(tetradecanyl)amino(C₂)alkylene and an ammonium salt thereof wherein;

R⁵ and R⁶ are both methyl; and

R⁷ is (C₁₄)alkyl.

9. The compound of claim 5, wherein:

R¹ is (C₁₄)acyloxy(C₁₃)alkyl.

10. The compound of claim 5, wherein:

R⁴ is a member selected from the group consisting of (C₁₅)acyloxy(C₂)alkyleneamino(C₂)alkylene; and

an ammonium salt wherein;

R⁷ is (C₁₅)acyloxy(C₂)alkylene.

11. The compound of claim 3, wherein:

said compound is N,N-dimethyl-N-tetradecyl-N-(2-[3-(3,5,8-trioxa-4-tridecylbicyclo[2.2.2]octyl)propanoyloxy]ethyl) ammonium iodide.

12. The compound of claim 3, wherein:

said compound is (N,N-dimethyl-N-(2-[3-(3,5,8-trioxa-4-heptylbicyclo[2.2.2]octyl)propanoyloxy]ethyl)-N-(2-tetradecanoyloxy)ethyl ammonium iodide.

13. A lipid formulation comprising a compound of the formula

wherein:

R¹ is a member selected from the group consisting of optionally substituted (C₇-C₁₇)alkyl, optionally substituted (C₇-C₁₇)alkenyl and optionally substituted (C₇-C₁₇)alkynyl;

R² is a member selected from the group consisting of (C₁-C₁₈)alkoxy and (C₁-C₁₈)alkylthio;

R³ is hydrogen;

alternatively,

R² and R³ and the carbons to which they are bound join to form a 5,6-membered;

a 6,6-membered;

a 6,7-membered;

or a 7,7-membered bicyclic ortho ester or ortho thioester ring;

A and A¹ are members independently selected from the group consisting of oxygen and sulfur;

x and y are independently selected from the integers 0, 1 and 2;

Z is a member selected from the group consisting of optionally substituted alkylene, optionally substituted alkyleneoxyalkylene and optionally substituted alkyleneaminoalkylene;

Q is a member selected from the group consisting of carboxyl, thiocarboxyl, dithiocarboxyl, carbonate, carbamate, phospho, phosphothio, phosphoro and thiophosphoro; and

R⁴ is a nitrogen containing headgroup wherein the nitrogen can be unsubstituted, mono-substituted, di-substituted, or a quaternary nitrogen salt and wherein the nitrogen substituent(s) are member(s) independently selected from the group consisting of optionally substituted (C₁-C₁₈)alkyl, optionally substituted (C₁-C₁₈)alkenyl, and optionally substituted (C₁-C₁₈)alkynyl and wherein R⁴ and Q are optionally linked with a (C₁-C₅)alkylene or (C₂-C₅)alkenyl group; and

a bioactive agent.

14. The lipid formulation of claim 13, wherein said bioactive agent is a nucleic acid.

15. The lipid formulation of claim 13, wherein said compound is formulated into a liposome.

16. The lipid formulation of claim 13, wherein said compound has the formula:

wherein;

R¹ is a member selected from the group consisting of (C₇-C₁₇)alkyl and substituted (C₇-C₁₇)alkyl;

Z is optionally substituted alkylene;

Q is carboxyl;

R⁴ is a member selected from the group consisting of optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene and a quaternary nitrogen salt having the structure

17. The lipid formulation of claim 16, wherein:

R¹ is (C₇-C₁₇)alkyl optionally substituted with one or more members selected from the group consisting of lower alkyl, acyl, halogen, hydroxy, amino, alkoxy, alkylamino, acylamino, acyloxy, and mercapto;

Z is (C₁-C₃) alkylene optionally substituted with one or more members selected from the group consisting of lower alkyl, aryl, acyl, halogen, hydroxy, amino, alkoxy, alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, mercapto, both saturated and unsaturated cyclic hydrocarbons and heterocycles;

R⁴ is a member selected from the group consisting of optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene or optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene, wherein the substituent is one or more members selected from the group consisting of lower alkyl, aryl, acyl, halogen, hydroxy, amino, alkoxy, alkylamino, acylamino, acyloxy, aryloxy, aryloxyalkyl, mercapto, both saturated and unsaturated cyclic hydrocarbons and heterocycles; and

an ammonium salt having the structure;

18. The lipid formulation of claim 17, wherein:

R¹ is (C₇-C₁₇)alkyl optionally substituted with (C₇-C₁₈)acyloxy;

Z is (C₁-C₃) alkylene optionally substituted with (C₇-C₁₈)alkoxy; and

R⁴ is a member selected from the group consisting of optionally substituted (C₁-C₁₈)alkylamino(C₁-C₅)alkylene, optionally substituted (C₁-C₁₈)dialkylamino(C₁-C₅)alkylene wherein the substituent is (C₁-C₁₈)acyloxy, and an ammonium salt wherein;

R⁶ is methyl.

19. The lipid formulation of claim 17, wherein:

R¹ is a (C₁₄-C₁₇)alkyl;

Z is (C₁-C₃)alkylene;

R⁴ is a member selected from the group consisting of (C₁₄-C₁₈)acyloxy(C₁-C₅)alkylamino(C₂-C₄)alkylene and an ammonium salt wherein;

R⁶ is methyl; and

R⁷ is (C₁₄-C₁₈)acyloxy(C₁-C₅)alkylene.

20. The lipid formulation of claim 17, wherein:

R¹ is (C₁₃)alkyl;

Z is (C₂)alkylene; and

R⁴ is a member selected from the group consisting of N-methyl-N-(tetradecanyl)amino(C₂)alkylene and an ammonium salt thereof wherein;

R⁶ is methyl; and

R⁷ is (C₁₄)alkyl.

21. The lipid formulation of claim 17, wherein:

R¹ is (C₁₄)acyloxy(C₁₃)alkyl.

22. The lipid formulation of claim 17, wherein:

R⁴ is a member selected from the group consisting of (C₁₅)acyloxy(C₂)alkyleneamino(C₂)alkylene; and

an ammonium salt wherein;

R⁷ is (C₁₅)acyloxy(C₂)alkylene.

23. A method for transfecting a cell, said method comprising:

contacting said cell with a lipid formulation comprising a compound of the formula

wherein;

R¹ is a member selected from the group consisting of optionally substituted (C₇-C₁₇)alkyl, optionally substituted (C₇-C₁₇)alkenyl and optionally substituted (C₇-C₁₇)alkynyl;

R² is a member selected from the group consisting of (C₁-C₁₈)alkoxy and (C₁-C₁₈)alkylthio;

R³ is hydrogen;

alternatively,

R² and R³ and the carbons to which they are bound join to form a 5,6-membered;

a 6,6-membered;

a 6,7-membered;

or a 7,7-membered bicyclic ortho ester or ortho thioester ring;

A and A¹ are members independently selected from the group consisting of oxygen and sulfur;

x and y are independently selected from the integers 0, 1 and 2;

Z is a member selected from the group consisting of optionally substituted alkylene, optionally substituted alkyleneoxyalkylene and optionally substituted alkyleneaminoalkylene;

Q is a member selected from the group consisting of carboxyl, thiocarboxyl, dithiocarboxyl, carbonate, carbamate, phospho, phosphothio, phosphoro and thiophosphoro; and

R⁴ is a nitrogen containing headgroup wherein the nitrogen can be unsubstituted, mono-substituted, di-substituted, or a quaternary nitrogen salt and wherein the nitrogen substituent(s) are member(s) independently selected from the group consisting of optionally substituted (C₁-C₁₈)alkyl, optionally substituted (C₁-C₁₈)alkenyl, and optionally substituted (C₁-C₁₈)alkynyl and wherein R⁴ and Q are optionally linked with a (C₁-C₅)alkylene or (C₂-C₅)alkenyl group; and

a nucleic acid, thereby transfecting said cell.

24. The method of claim 23, wherein said lipid formulation comprises a liposome.

25. A method for delivering a bioactive agent to a cell, said method comprising:

contacting said cell with a lipid formulation comprising a compound of the formula

wherein;

R¹ is a member selected from the group consisting of optionally substituted (C₇-C₁₇)alkyl, optionally substituted (C₇-C₁₇)alkenyl and optionally substituted (C₇-C₁₇)alkynyl;

R² is a member selected from the group consisting of (C₁-C₁₈)alkoxy and (C₁-C₁₈)alkylthio;

R³ is hydrogen;

alternatively,

R² and R³ and the carbons to which they are bound join to form a 5,6-membered;

a 6,6-membered;

a 6,7-membered;

or a 7,7-membered bicyclic ortho ester or ortho thioester ring;

A and A¹ are members independently selected from the group consisting of oxygen and sulfur;

x and y are independently selected from the integers 0, 1 and 2;

Z is a member selected from the group consisting of optionally substituted alkylene, optionally substituted alkyleneoxyalkylene and optionally substituted alkyleneaminoalkylene;

Q is a member selected from the group consisting of carboxyl, thiocarboxyl, dithiocarboxyl, carbonate, carbamate, phospho, phosphothio, phosphoro and thiophosphoro; and

R⁴ is a nitrogen containing headgroup wherein the nitrogen can be unsubstituted, mono-substituted, di-substituted, or a quaternary nitrogen salt and wherein the nitrogen substituent(s) are member(s) independently selected from the group consisting of optionally substituted (C₁-C₁₈)alkyl, optionally substituted (C₁-C₁₈)alkenyl, and optionally substituted (C₁-C₁₈)alkynyl and wherein R⁴ and Q are optionally linked with a (C₁-C₅)alkylene or (C₂-C₅)alkenyl group; and

a bioactive agent,

thereby delivering said bioactive agent to said cell.

26. The method of claim 25, wherein said bioactive agent is a member selected from the group consisting of antimicrobials, antibiotics, antimyobacterials, antifungals, antivirals, neoplastic agents, agents affecting the immune response, blood calcium regulators, agents useful in glucose regulation, anticoagulants, antithrombotics, antihyperlipidemic agents, cardiac drugs, thyromimetic drugs, antithyroid drugs, adrenergics, antihypertensive agents, cholinergics, anticholinergics, antispasmodics, antiulcer agents, skeletal and smooth muscle relaxants, prostaglandins, general inhibitors of the allergic response, antihistamines, local anesthetics, analgesics, narcotic antagonists, antitussives, sedative-hypnotic agents, anticonvulsants, antipsychotics, anti-anxiety agents, antidepressant agents, anorexigenics, non-steroidal anti-inflammatory agents, steroidal anti-inflammatory agents, antioxidants, vaso-active agents, bone-active agents, antiarthritics, diagnostic agents, antineoplastic agents and anti-infective agents.

27. The method of claim 25, wherein said bioactive agent is a small molecule.

28. The method of claim 25, wherein said lipid formulation is a liposome.