阅读说明：本技术 脂质体药物递送媒介物 (Liposomal drug delivery vehicles ) 是由 J.克莱恩 R.戈德伯格 W.林 于 2018-02-16 设计创作，主要内容包括：在此公开了用于将治疗活性剂递送至需要其的受试者的脂质体。该脂质体包含：a)至少一种形成双层的脂质；b)具有通式I的聚合化合物：(I)其中m、n、L、X、Y和Z如本文所定义；和c)包含在脂质体中和/或脂质体表面上的治疗活性剂。<Image he="115" wi="115" file="100004_DEST_PATH_IMAGE002.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>式I。(Disclosed herein are liposomes for delivering a therapeutically active agent to a subject in need thereof. The liposome comprises: a) at least one bilayer-forming lipid; b) a polymeric compound having the general formula I: (I) wherein m, n, L, X, Y and Z are as defined herein; and c) a therapeutically active agent contained in and/or on the surface of the liposomes. Formula I.)

1. Liposomes comprising

a) At least one bilayer-forming lipid;

b) a polymeric compound having the general formula I:

formula I

Wherein:

m is zero or a positive integer;

n is an integer of at least 1, wherein n is at least 2 when X does not comprise a phosphate group;

x is a lipid moiety;

y is a backbone unit forming a polymeric backbone;

l is absent or a linking moiety; and

z has the general formula II:

formula II

Wherein:

a is a substituted or unsubstituted hydrocarbon;

b is an oxygen atom or is absent; and

R₁-R₃each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, heteroalicyclic, aryl, and heteroaryl; and

c) a therapeutically active agent contained in and/or on the surface of the liposomes,

the liposomes are useful for delivering the therapeutically active agent to a subject in need thereof.

2. A liposome for use according to claim 1, for use in the treatment of a medical condition treatable by the therapeutically active agent in the subject.

3. The liposome for use according to any of claims 1 to 2, wherein said delivery comprises sustained release of said therapeutically active agent.

4. The liposome for use according to any one of claims 1 to 3, wherein the therapeutically effective agent is selected from the group consisting of analgesics, anti-inflammatory agents, antiproliferative agents, antimicrobial agents and vaccine antigens.

5. The liposome for use according to any of claims 1 to 4, wherein the delivery is achieved by parenteral systemic administration.

6. The liposome for use according to any of claims 1 to 4, wherein the delivery is achieved by intra-articular administration.

7. The liposome for use according to any one of claims 1 to 6, for use in the treatment of synovial joint disorders.

8. The liposome for use according to claim 7, wherein the synovial joint disorder is selected from the group consisting of arthritis, bursitis, carpal tunnel syndrome, fibromyositis, gout, locked joints, tendonitis, traumatic joint injury and joint injury associated with surgery.

9. The liposome for use according to any of claims 1 to 8, wherein the therapeutically active agent is an analgesic and/or an anti-inflammatory agent.

10. Liposome for use according to any of claims 1 to 9, wherein the molar ratio of the bilayer-forming lipid and the polymeric compound is in the range of 5:1 to 5000: 1, in the above range.

11. The liposome for use according to any of claims 1 to 10, wherein Y is a substituted or unsubstituted alkylene unit.

12. The liposome for use according to claim 11, wherein Y is a substituted or unsubstituted ethylene unit.

13. A liposome for use according to claim 12, wherein Y has the formula-CR₄R₅-CR₆D-, wherein:

d is R when Y is a backbone unit not linked to said L or said Z₇(ii) a And when Y is a backbone unit attached to said L or said Z, D is a covalent bond or a linking group linking Y to said L or said Z, the linking group being selected from the group consisting of-O-, -S-, alkylene, arylene, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido and amino; and

R₄-R₇each independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, azide, azo, phosphate, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido, and amino.

14. The liposome for use according to claim 13, wherein R₄And R₅Each is hydrogen.

15. The liposome for use according to any one of claims 13 to 14, wherein R₆Is hydrogen.

16. The liposome for use according to any one of claims 13 to 15, wherein the linker is selected from the group consisting of-O-, -C (= O) NH-, and phenylene.

17. The liposome for use according to claim 16, wherein the linking group is-C (= O) O-.

18. The liposome for use according to any of claims 13 to 17, wherein L is a substituted or unsubstituted hydrocarbon of 1 to 10 carbon atoms in length.

19. The liposome for use according to claim 18, wherein L is a substituted or unsubstituted ethylene group.

20. The liposome for use according to any one of claims 1 to 19, wherein B is an oxygen atom.

21. The liposome for use according to any of claims 1 to 20, wherein a is a substituted or unsubstituted hydrocarbon of 1 to 4 carbon atoms in length.

22. The liposome for use according to claim 21, wherein a is a substituted or unsubstituted ethylene group.

23. The liposome for use according to any one of claims 1 to 22, wherein R₁-R₃Each independently is hydrogen or C_1-4-an alkyl group.

24. The liposome for use according to claim 23, wherein R₁-R₃Each is methyl.

25. The liposome for use according to any one of claims 1 to 24, wherein n is at least 3.

26. The liposome for use according to claim 25, wherein n is in the range of 5 to 50 and m is in the range of 0 to 50.

27. The liposome for use according to any of claims 1 to 26, wherein at least a part of said Y, said L and/or said Z comprises at least one targeting moiety.

28. The liposome for use according to claim 27, wherein the polymeric compound has the general formula Ib:

formula Ib

Wherein:

t is the unit of Y comprising the at least one targeting moiety;

x and T are attached to the distal end of the polymeric compound; and

x, Y, L, Z, n and m are as defined for formula I, provided that m is a positive integer.

29. The liposome for use according to any of claims 1 to 28, wherein the lipid is selected from the group consisting of fatty acids, monoglycerides, diglycerides, triglycerides, glycerophospholipids, sphingolipids and sterols.

30. The liposome for use according to claim 29, wherein the glycerophospholipid is selected from the group consisting of phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol and phosphatidylinositol.

31. The liposome for use according to any one of claims 1 to 28, wherein X has the general formula III:

formula III

Wherein:

W₁and W₂Each independently selected from hydrogen, alkyl, alkenyl, alkynyl and acyl, wherein W is₁And W₂Is not hydrogen;

j is-P (= O) (OH) -O-or absent;

k is a substituted or unsubstituted hydrocarbon of 1 to 10 carbon atoms in length, or is absent;

m is a linking group selected from-O-, -S-, amino, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea, thiourea, carbamoyl, thiocarbamoyl, amido, carboxyl and sulfonamide, or is absent; and

q is a substituted or unsubstituted hydrocarbon of 1 to 10 carbon atoms in length,

wherein when M is absent, K is also absent.

32. The liposome for use according to claim 31, wherein J is-P (= O) (OH) -O-and K is selected from an ethanolamine moiety, a serine moiety, a glycerol moiety, and an inositol moiety.

33. A liposome for use according to any one of claims 31 to 32 wherein M is amido.

34. The liposome for use according to claim 31, wherein J and K are absent and M is a carbonyl group.

35. The liposome for use according to any one of claims 31 to 34, wherein Q is dimethylmethylene (-C (CH)₃)₂-)。

36. The liposome for use according to any one of claims 31 to 35, wherein W is₁And W₂Is an alkyl, alkenyl, alkynyl or acyl group of 10 to 30 carbon atoms in length.

37. Liposome for use according to any of claims 1 to 36, wherein the lipid fraction comprises at least one fatty acid moiety selected from the group consisting of lauroyl, myristoyl, palmitoyl, stearoyl, palmitoyl, oleoyl and linoleoyl.

38. The liposome for use according to any one of claims 1 to 37, formulated as part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

39. The liposome for use according to claim 38, wherein the carrier comprises an aqueous liquid.

40. The liposome for use according to claim 38 or 39, wherein the pharmaceutical composition further comprises a water-soluble biopolymer.

41. The liposome for use according to claim 40, wherein the biopolymer comprises hyaluronic acid.

Field and background of the invention

The present invention, in some embodiments thereof, relates to therapeutics, and more particularly, but not exclusively, to novel polymeric compounds that are particularly useful in forming drug delivery vehicles.

Liposomes have been extensively studied as drug carriers because they can contain essentially any kind of drug-including both hydrophilic and hydrophobic molecules. However, standard liposomes are rapidly removed from the blood mainly by cells of the mononuclear phagocyte system [ Blume& Cevc, Biochim Biophys Acta 1990, 1029, 91-97]。

It has been reported that the use of phosphatidylethanolamine liposomes with polyethylene glycol covalently attached to phosphatidylethanolamine (PE-PEG) results in long circulation times and high retention of pegylated liposomes compared to pure (non-pegylated) liposomes [ Blume& Cevc, Biochim Biophys Acta - Biomembranes 1990, 1029, 91-97]. Pegylated liposomes are used in doxorubicin formulations to treat cancer (e.g., Caelyx, Doxil [ [ Charrois ] Tokys [) ® cancers& Allen,Biochim Biophys Acta- Biomembranes 2004, 1663, 167-177]。

Osteoarthritis (OA) is a degenerative joint disease with progressive loss of articular cartilage, subchondral sclerosis, osteophyte formation, synovial changes and increased synovial fluid mass with reduced viscosity and lubrication properties [ Gerwin et al,Adv Drug Deliv Rev 2006, 58:226-242]. OA is the most common type of arthritis causing a significant burden [ Hunter et al,Nat Rev Rheumatol 2014, 10:437-441]and affects a large number of people: for example, there are approximately 3000 million people in the United states, 4000 million people in Europe, and eight hundred and fifty million people in the United kingdom, with six million people suffering from sustained pain [ Hurley ]& Carter,Perspect Public Health 2016, 136:67-69]。

The local administration of drugs to treat OA by intra-articular (IA) injection may reduce systemic exposure to the drugs and improve the delivery efficiency of the target area, while allowing for increased levels of the therapeutic agents. Various glucocorticoid and Hyaluronic Acid (HA) formulations are available for IA treatment, providing at best only short-term pain relief. Of the drug after IA injectionThe half-life is several hours. This short half-life requires repeated injections, which results in poor patient compliance and a high risk of introducing bacteria into the joint space [ Butoescu et al,Eur J Pharm Biopharm 2009, 73:2015-218]。

it has been reported that the administration of liposome-embedded methotrexate by IA injection for the treatment of antigen-induced arthritis in rabbits resulted in improved efficacy relative to the injection of free drug. However, this enhanced effect was limited to inhibition of arthritis after 7 days rather than inhibition of arthritis after 21 days [ Foong& Green, J Pharm Pharmacol 1993, 45:204-209]. Diclofenac sodium loaded into a liposome matrix for IA administration has been reported to inhibit joint swelling in antigen-induced arthritis [ Turker et al,J Drug Target 2008, 16:51-57]. Dexamethasone palmitate encapsulated in lipid microspheres was reported to show enhanced retention and sustained therapeutic effect [ Labrenz ]& Rose, Clin Drug Invest2001, 21:429-436; Bonanomi et al.,Rheumatol Int 1987, 7:203-212]. Liposomal dexamethasone palmitate, commercially available in germany under the trade name Lipotalon @ (Merckle), is the only commercially available liposomal formulation for IA delivery, although several liposomal formulations for IA delivery have entered clinical trials [ Evans et al,Nat Rev Rheumatol 2014, 10:11-22]。

intra-articular delivery of liposomal formulations has irreproducibility problems associated with the polydispersity and aggregation of liposomes composed of multilamellar vesicle (MLV) vehicles, or the problem of rapid clearance in the case of Smaller Unilamellar Vesicles (SUV) [ bonomi et al,Rheumatol Int 1987, 7:203-212]. Liposomal aggregates injected into the body are more prone to protein adsorption and are attacked and removed by macrophages [ Moghimi& Szebeni, Prog Lipid Res 2003, 42:463-478]。

Other methods of arthritis treatment include IA injection of a viscous supplement of hyaluronic acid [ Evans et al,Nat Rev Rheumatol 2014, 10:11-22]and the use of liposomes as a biolubricant [ Sivan et al,Langmuir 2010, 26:1107-1116]。

U.S. patent No. 8,617,592 describes block copolymers and conjugates comprising zwitterionic poly (carboxybetaine), poly (sulfobetaine), or poly (phosphobetaine) blocks and hydrophobic blocks, which self-assemble into particles, and the use of such particles for the delivery of therapeutic and diagnostic agents.

International patent application publication No. WO2017/109784 describes polymeric compounds comprising a lipid moiety and an ionic polymeric moiety, such as a pMPC (poly (O- (2-methacryloyloxyethyl) phosphorylcholine)) moiety, and bilayers and liposomes comprising a combination of such polymeric compounds with lipids forming the bilayer. Bilayers and/or liposomes comprising such polymeric compounds are described as being useful for reducing the coefficient of friction of a surface and/or for inhibiting biofilm formation.

Other background art includes Chen et alScience 2009, 323:1698-1702]; Gabizon et al. [Cancer Res 1994, 54:987-982]; Goldberg et al. [Biophys J 2011, 100:2403-2411]; Goldberg et al. [Adv Materials 2011, 23:3517-3521]; Goldberg & Klein [Chem Phys Lipids 2012, 165:374-381]; Harris & Chess [Nat Rev Drug Discov2003, 2:214-221]; Seror et al. [Nat Commun 2015, 6:6497]; Sorkin et al. [Biomaterials 2014, 34:5465-5475]International patent application Nos. PCT/IL2014/050604 (published as WO 2015/001564), PCT/IL2015/050605 (published as WO 2015/193887) and PCT/IL2015/050606 (published as WO2015/193888), and Israel patent application No. 234929.

Summary of The Invention

The present inventors have now surprisingly found that recently designed lipid-derived polymeric compounds, in which the monomers comprise both phosphate and ammonium ionic groups, can be effectively used to deliver therapeutically active agents to a body site of a subject.

Embodiments of the invention relate to the use of these compounds for drug delivery (e.g., sustained release of a drug), thereby treating a medical condition treatable by the drug.

According to an aspect of some embodiments of the present invention there is provided a liposome comprising:

a) at least one bilayer-forming lipid;

b) a polymeric compound having the general formula I:

formula I

Wherein:

m is zero or a positive integer;

n is an integer of at least 1, wherein n is at least 2 when X does not comprise a phosphate group;

x is a lipid moiety;

y is a backbone unit forming a polymeric backbone;

l is absent or a linking moiety; and

z has the general formula II:

formula II

Wherein:

a is a substituted or unsubstituted hydrocarbon;

b is an oxygen atom or is absent; and

R₁-R₃each independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, heteroalicyclic, aryl, and heteroaryl; and

c) a therapeutically active agent contained in and/or on the surface of the liposomes,

the liposomes are useful for delivering a therapeutically active agent to a subject in need thereof.

According to some embodiments of the invention, the liposome is for use in treating a medical condition treatable by a therapeutically active agent in a subject.

According to some embodiments of the invention, the delivery comprises sustained release of the therapeutically active agent.

According to some embodiments of the invention, the therapeutically effective agent is selected from the group consisting of analgesics, anti-inflammatory agents, antiproliferative agents, antimicrobial agents, and vaccine antigens.

According to some embodiments of the invention, the delivery is achieved by parenteral systemic administration.

According to some embodiments of the invention, the delivery is achieved by intra-articular administration.

According to some embodiments of the invention, the liposome is for use in treating a synovial joint disorder.

According to some embodiments of the invention, the synovial joint disorder is selected from the group consisting of arthritis, bursitis, carpal tunnel syndrome, fibromyositis, gout, locked joints, tendonitis, traumatic joint injury, and joint injury associated with surgery.

According to some embodiments of the invention, the therapeutically active agent is an analgesic and/or an anti-inflammatory agent.

According to some embodiments of the invention, the lipid and the polymeric compound forming the bilayer are present in a molar ratio of 5:1 to 5000: 1, in the above range.

According to some embodiments of the invention, Y is a substituted or unsubstituted alkylene unit.

According to some embodiments of the invention, Y is a substituted or unsubstituted ethylene unit.

According to some embodiments of the invention, Y has the formula-CR₄R₅-CR₆D-, wherein:

when Y is a backbone unit not linked to L or Z, D is R₇(ii) a And when Y is a backbone unit attached to L or Z, D is a covalent bond or a linking group linking Y to L or Z selected from the group consisting of-O-, -S-, alkylene, arylene, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido, and amino; and

R₄-R₇each independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, azide, azo, phosphate, phosphonyl, phosphinyl, oxyAnd (b) a substituent selected from the group consisting of substituted, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido and amino.

According to some embodiments of the invention, R₄And R₅Each is hydrogen.

According to some embodiments of the invention, R₆Is hydrogen.

According to some embodiments of the invention, the linking group is selected from the group consisting of-O-, -C (= O) NH-, and phenylene.

According to some embodiments of the invention, the linking group is-C (= O) O-.

According to some embodiments of the invention, L is a substituted or unsubstituted hydrocarbon from 1 to 10 carbon atoms in length.

According to some embodiments of the invention, L is substituted or unsubstituted ethylene.

According to some embodiments of the invention, B is an oxygen atom.

According to some embodiments of the invention, a is a substituted or unsubstituted hydrocarbon of 1 to 4 carbon atoms in length.

According to some embodiments of the invention, a is substituted or unsubstituted ethylene.

According to some embodiments of the invention, R₁-R₃Each independently is hydrogen or C_1-4-an alkyl group.

According to some embodiments of the invention, R₁-R₃Each is methyl.

According to some embodiments of the invention, n is at least 3.

According to some embodiments of the invention, n is in the range of 5 to 50, and m is in the range of 0 to 50.

According to some embodiments of the invention, at least a portion of Y, L and/or Z comprises at least one targeting moiety.

According to some embodiments of the invention, the polymeric compound has the general formula Ib:

formula Ib

Wherein:

t is a unit of Y comprising the at least one targeting moiety;

x and T are attached to the distal end of the polymeric compound; and

x, Y, L, Z, n and m are as defined for formula I, provided that m is a positive integer.

According to some embodiments of the invention, the lipid is selected from the group consisting of fatty acids, monoglycerides, diglycerides, triglycerides, glycerophospholipids, sphingolipids and sterols.

According to some embodiments of the invention, the glycerophospholipid is selected from the group consisting of phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol and phosphatidylinositol.

According to some embodiments of the invention, X has the general formula III:

formula III

Wherein:

W₁and W₂Each independently selected from hydrogen, alkyl, alkenyl, alkynyl and acyl, wherein W is₁And W₂Is not hydrogen;

j is-P (= O) (OH) -O-or absent;

k is a substituted or unsubstituted hydrocarbon of 1 to 10 carbon atoms in length, or is absent;

m is a linking group selected from-O-, -S-, amino, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea, thiourea, carbamoyl, thiocarbamoyl, amido, carboxyl and sulfonamide, or is absent; and

q is a substituted or unsubstituted hydrocarbon of 1 to 10 carbon atoms in length,

wherein when M is absent, K is also absent.

According to some embodiments of the invention, J is-P (= O) (OH) -O-and K is selected from an ethanolamine moiety, a serine moiety, a glycerol moiety, and an inositol moiety.

According to some embodiments of the invention, M is amido.

According to some embodiments of the invention, J and K are absent and M is carbonyl.

According to some embodiments of the invention, Q is dimethylmethylene (-C (CH)₃)₂-)。

According to some embodiments of the invention, W₁And W₂Is an alkyl, alkenyl, alkynyl or acyl group of 10 to 30 carbon atoms in length.

According to some embodiments of the invention, the lipid fraction comprises at least one fatty acid moiety selected from lauroyl, myristoyl, palmitoyl, stearoyl, palmitoyl, oleoyl and linoleoyl.

According to some embodiments of the invention, the liposome is formulated as part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.

According to some embodiments of the invention, the carrier comprises an aqueous liquid.

According to some embodiments of the invention, the pharmaceutical composition further comprises a water-soluble biopolymer.

According to some embodiments of the invention, the biopolymer comprises hyaluronic acid.

Unless defined otherwise, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be necessarily limiting.

Brief description of several views of the drawings

Some embodiments of the invention are described herein, by way of example only, with reference to the accompanying drawings. Referring now in detail to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of the embodiments of the present invention. In this regard, the description taken with the drawings make it apparent to those skilled in the art how the embodiments of the invention may be practiced.

In the drawings:

figure 1 presents images showing the fluorescence intensity of mice 147 hours after intraarticular injection of fluorescently labeled hyaluronic acid (left mice) or fluorescently labeled hydrogenated soy phosphatidylcholine liposomes with a pMPC moiety, according to some embodiments of the present invention; labeled with the fluorescent dye DiR.

FIGS. 2A-2D present curves showing the fluorescence signal in mice as a function of time after intraarticular injection of fluorescently labeled hyaluronic acid (HA; FIG. 2A) or fluorescently labeled hydrogenated soy phosphatidylcholine liposomes having a 2kDa pMPC moiety (liposome-pMPC in FIG. 2A; HSPC-pMPC in FIG. 2B), a 2kDa pMPC and a trimethylammonium propane moiety (liposome-pMPC/TAP in FIG. 2A; HSPC-TAP/pMPC in FIG. 2D) or a 2kDa PEG moiety (liposome-PEG in FIG. 2A; HSPC-PEG in FIG. 2C); the exponential fit and the associated calculated half-lives (T1/2) of 95, 22 and 90 hours are shown in FIGS. 2B-2D, respectively; the mean liposome diameter was 170nm or 162 nm.

Figure 3 presents a curve showing the fluorescence signal in mice as a function of time following intra-articular injection of fluorescently labeled Hydrogenated Soybean Phosphatidylcholine (HSPC) liposomes with a 2kDa PEG moiety, and an exponential fit of the data (from which a calculated half-life of 26 hours was derived); the mean liposome diameter was 170 nm.

Figure 4 presents curves showing the fluorescence signal in mice as a function of time after intraarticular injection of fluorescently labeled liposomes having an average diameter of 80nm comprising Hydrogenated Soy Phosphatidylcholine (HSPC) or HSPC with a 2kDa peg moiety or a 2kDa pMPC moiety, alone or in combination with Hyaluronic Acid (HA); also presented are calculated half-lives (T)_1/2) The mean liposome diameter was 80 nm.

Figure 5 presents a graph showing the fluorescence signal in mice as a function of time following intra-articular injection of fluorescently labeled Hydrogenated Soy Phosphatidylcholine (HSPC) liposomes with a 550Da PEG moiety, and an exponential fit of the data from which a calculated half-life of 10 hours was derived; the mean liposome diameter was 170 nm.

Description of specific embodiments of the invention

The present invention, in some embodiments thereof, relates to therapeutics, and more particularly, but not exclusively, to novel polymeric compounds that are particularly useful in forming drug delivery vehicles.

Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or illustrated by the examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

When studying liposomes in vivo, the inventors have occasionally found that liposomes stabilized with lipid-derived polymeric compounds, wherein the monomers comprise both phosphate and ammonium groups, have particularly long retention times in vivo. As demonstrated in the examples section below, the inventors have found that liposomes comprising such polymeric compounds are suitable for drug delivery and are advantageous compared to prior art pegylated liposomes for drug delivery, e.g. exhibit longer retention times than comparable pegylated liposomes.

Thus, embodiments of the present invention relate to the use of such liposomes as drug delivery vehicles, i.e. to liposomes comprising therapeutically active agents and their use in the treatment of medical conditions. In some embodiments, the liposomes are used to treat medical conditions where it is desirable or necessary to extend the retention time of the therapeutically active agent in the liposome.

According to an aspect of some embodiments of the present invention there is provided a liposome comprising at least one bilayer-forming lipid, a polymeric compound according to any of the respective embodiments described herein, and a therapeutically active agent contained in and/or on the liposome. In some embodiments, the liposomes are used to deliver a therapeutically active agent to a subject in need thereof.

According to an aspect of some embodiments of the present invention there is provided a use of a liposome comprising at least one bilayer-forming lipid, a polymeric compound according to any of the respective embodiments described herein, and a therapeutically active agent contained in and/or on the liposome, in the manufacture of a medicament for delivering the therapeutically active agent to a subject in need thereof.

According to an aspect of some embodiments of the present invention there is provided a method of delivering a therapeutically active agent to a subject in need thereof, the method comprising administering to the subject a liposome comprising at least one bilayer-forming lipid, a polymeric compound according to any of the respective embodiments described herein, and a therapeutically active agent contained in and/or on the liposome, thereby delivering the therapeutically active agent to the subject in need thereof.

In some embodiments according to any embodiment of any aspect described herein, the liposome and/or the method described herein is used for the treatment of a medical condition in a subject treatable by a therapeutically active agent (according to any respective embodiment described herein).

In some embodiments according to any embodiment of any aspect described herein, delivering the therapeutically active agent comprises sustained release of the therapeutically active agent according to any respective embodiment described herein.

In some embodiments according to any embodiment of any aspect described herein, the liposome is selected to be capable of sustained release of the therapeutically active agent according to any respective embodiment described herein.

As used herein, "delivery" or "delivering" of a therapeutically active agent or drug (these terms are used interchangeably herein) refers to administering a therapeutically active agent to a subject while controlling the duration and/or proportion of the therapeutically active agent at a desired body site, depending on the condition of the subject (e.g., the body site where the therapeutically active agent desirably exerts a therapeutic effect). Thus, the terms "delivery" and "delivering" (and grammatical variations thereof) include targeting a therapeutically active agent to a particular body site such that a higher proportion of the therapeutically active agent reaches the body site (e.g., using a suitable targeting moiety); and/or control the duration of presence of such therapeutically active agents in the body (e.g., in the blood) -e.g., by sustained release-which may be correlated to the duration of such therapeutically active agents at a desired body site (even if the body site is not specifically targeted).

As used herein, "sustained release" refers to a formulation of an agent that provides for gradual and/or delayed ("sustained") release of the agent (e.g., from a reservoir such as a liposome according to any of the respective embodiments described herein) resulting in the agent being present at a body site (e.g., in the blood upon systemic administration, or in a body site where the agent is administered locally) for a longer duration and/or at a later time (relative to administration) than if the agent itself were administered (by the same route of administration).

For example, in the context of embodiments of the present invention, an agent administered per se (other than a sustained release formulation) optionally refers to a formulation of the agent without liposomes according to embodiments of the present invention and comprising the same carrier (if any) as the sustained release formulation.

In some embodiments, the sustained release is characterized by a concentration of the therapeutically active agent (e.g., in the blood when administered systemically, or in a body site where the agent is administered locally) that is at least half the maximum concentration (Cmax) for a period of time that is at least 50% more than the corresponding period of time for the agent when administered by itself (e.g., as defined herein) in an amount that results in the same maximum concentration (i.e., a period of time for which the agent concentration is at least half the maximum concentration). In some such embodiments, the time period (for sustained release) is at least 100% (i.e., two times) more than the corresponding time period (for the agent itself). In some embodiments, the time period (for sustained release) is at least 200% (i.e., 3-fold) more than the corresponding time period (for the agent itself). In some embodiments, the time period (for sustained release) is at least 400% (i.e., 5-fold) more than the corresponding time period (for the agent itself).

In some embodiments, the sustained release is characterized by a concentration of the therapeutically active agent (e.g., in the blood when administered systemically, or in a body site where the agent is administered locally) that is at least half the maximum concentration (Cmax) for a period of at least 6 hours. In some such embodiments, the period of time is at least 12 hours. In some embodiments, the period of time is at least 24 hours. In some embodiments, the period of time is at least 2 days. In some embodiments, the period of time is at least 4 days. In some embodiments, the period of time is at least one week. In some embodiments, the period of time is at least 2 weeks. In some embodiments, the period of time is at least 4 weeks.

Sustained release (according to any of the respective embodiments described herein) may allow for a regimen characterized, for example, by a lower frequency of administration and/or a greater therapeutic effect for any given administration. Based on the duration of sustained release (e.g., the period of time during which the agent concentration is at least half the maximum concentration and/or the agent concentration is at least the minimum effective concentration according to any of the respective embodiments described herein), and the ratio between the desired maximum concentration and the minimum effective concentration of a given agent (e.g., the "therapeutic window" of the agent), the skilled artisan will be able to readily determine the appropriate frequency of administration of a given therapeutically active agent.

Polymeric compound (b):

according to some embodiments of any of the embodiments described herein, the liposome comprises a polymeric compound (according to any of the respective embodiments described herein) having the general formula I:

formula I

Wherein:

m is zero or a positive integer;

n is an integer of at least 1;

x is a lipid moiety, wherein n is at least 2 when X does not comprise a phosphate group;

y is a backbone unit forming a polymeric backbone;

l is absent or a linking moiety; and

z has the general formula II:

formula II

Wherein:

a is a substituted or unsubstituted hydrocarbon;

b is an oxygen atom or is absent; and

R₁-R₃each independently hydrogen, alkyl, cycloalkyl, heteroalicyclic, aryl or heteroaryl,

as described in more detail below.

Formula I can also be described herein simply as:

X-[Y(-L-Z)]n[Y]m

which are considered interchangeable with the schematic description above.

Herein, the term "polymer" refers to a compound having at least 2 repeating units (and more preferably at least 3 repeating units), which are the same or similar. It will be appreciated that when n is at least 2, the compound of formula I is polymeric by definition in that it comprises at least 2 backbone units represented by Y.

Herein, the phrase "polymeric moiety" refers to a portion of a polymeric compound having the general formula Ia (according to any of the embodiments described herein relating to the general formula I):

formula Ia

Wherein m, n, Y, L and Z are as defined herein for formula I.

Formula Ia can also be described briefly herein as:

[Y(-L-Z)]n[Y]m

which are considered interchangeable with the schematic description above.

Herein, the phrase "polymeric compound" also includes compounds having a "polymeric moiety" as described herein, having one unit (e.g., according to formula Ia, wherein n is 1), provided that the lipid moiety described herein (e.g., the lipid moiety represented by X) has a similar unit. For example, when the lipid moiety comprises a phosphate group (e.g., the lipid moiety is a glycerophospholipid moiety) such that the lipid moiety has a phosphate group and a single unit of the polymeric moiety has a phosphate group, two phosphate groups may be considered to be repeating units.

However, in a preferred embodiment, n is at least 2, such that the polymeric moiety itself has at least two units. In some embodiments, n is at least 3.

As used herein, the term "backbone unit" refers to a repeating unit, wherein the linkage (e.g., sequential linkage) of a plurality of repeating units forms a polymeric backbone. The multiple linked repeat units themselves are also referred to herein as "polymeric backbones".

As shown in formulas I and Ia, L and Z together form a pendant group that is at least a portion of a backbone unit, which group is referred to herein simply as a "pendant group" for brevity.

Each backbone unit Y having a pendant group (i.e., the unit represented by Y (-L-Z), the number of which is represented by the variable n) and each backbone unit Y having no pendant group (the number of which is represented by the variable m) are also referred to herein as "monomer units".

The backbone unit can optionally be the residue of a polymerizable monomer or polymerizable portion of a monomer. Various polymerizable monomers and moieties will be known to those skilled in the art, and the structures (e.g., monomer units) of the residues of these monomers that are produced upon polymerization are also known to those skilled in the art.

"residue of a polymerizable monomer" refers to a modified form of the polymerizable monomer and/or a portion of the polymerizable monomer that remains after polymerization.

A portion of the polymerizable monomer can be formed, for example, by a condensation reaction, e.g., wherein at least one atom or group (e.g., a hydrogen atom or a hydroxyl group) in the monomer, and optionally at least two atoms or groups (e.g., a hydrogen atom and a hydroxyl group) in the monomer, are replaced by a covalent bond with another polymerizable monomer.

The polymerizable monomer in modified form can be formed, for example, by ring opening (wherein the covalent bond between two atoms in the ring is broken and the two atoms are optionally each attached to another polymerizable monomer); and/or by addition to an unsaturated bond, wherein the unsaturated bond between two adjacent atoms is broken (e.g., an unsaturated double bond is converted to a saturated bond, or an unsaturated triple bond is converted to an unsaturated double bond) and the two atoms are optionally each attached to another polymerizable monomer.

The polymerizable monomer in modified form may consist essentially of the same atoms as the original monomer, e.g., differ only in the rearrangement of covalent bonds, or may have a different atomic composition, e.g., where polymerization involves a condensation reaction (e.g., as described herein).

Examples of backbone units include, but are not limited to, substituted or unsubstituted hydrocarbons (which may form a substituted or unsubstituted hydrocarbon backbone), such as alkylene units; hydroxycarboxylic acid units (which may form a polyester backbone), such as glycolate, lactate, hydroxybutyrate, hydroxyvalerate, hydroxyhexanoate, and hydroxybenzoate units; dicarboxylic acid units (which may be combined with diols to form polyester backbones and/or diamines to form polyamide backbones), such as adipate, succinate, terephthalate, and naphthalate units; diol units (which may form a polyether backbone, or in combination with dicarboxylic acids form a polyester backbone), such as ethylene glycol, 1, 2-propanediol, 1, 3-propanediol, 1, 4-butanediol, and bisphenol a units; diamine units (which may be combined with dicarboxylic acids to form a polyamide backbone), for example p-phenylenediamine and alkylenediamines such as hexamethylenediamine; urethane units (which may form a polyurethane backbone); amino acid residues (which may form the polypeptide backbone); and sugar residues (which may form a polysaccharide backbone).

In some embodiments of any of the embodiments described herein, Y is a substituted or unsubstituted alkylene unit.

In some embodiments, Y is a substituted or unsubstituted ethylene unit, i.e., an alkylene unit 2 atoms in length.

The polymeric backbone in which Y is a substituted or unsubstituted ethylene unit may optionally be polymerized, for example, by polymerizing ethylene (CH)₂=CH₂) And/or substituted derivatives thereof (also referred to herein as "vinyl monomers"). This polymerization is a very well studied procedure and one of ordinary skill in the art will know of many techniques to achieve this polymerization.

It should be understood that any embodiment described herein that relates to a polymeric backbone formed by polymerization includes any polymeric backbone having a structure that can be formed by such polymerization, regardless of whether the polymeric backbone is formed by such polymerization (or any other type of polymerization) in practice.

As is well known in the art, the unsaturated bonds of ethylene and substituted ethylene derivatives become saturated upon polymerization, such that backbone units in the polymeric backbone are saturated, although they may be referred to as units of unsaturated compounds similar to them (e.g., "vinyl monomers" or "olefin monomers").

Polymers that can be formed from unsaturated monomers such as vinyl monomers and olefin monomers are also referred to by the terms "polyethylene" and "polyolefin".

Herein, "unsubstituted" alkylene units (e.g., ethylene units) refer to alkylene units that do not have any substituents other than the pendent groups discussed herein (denoted as (-L-Z)). That is, an alkylene unit attached to a pendant group described above is considered unsubstituted if there is no substituent at any other position on the alkylene unit.

In some of any of the embodiments described herein, Y has the formula-CR₄R₅-CR₆D-。

When Y is a backbone unit not linked to L or Z (i.e., to a pendant group as described herein), D is R₇(end groups, as defined herein); when Y is a backbone unit linked to L or Z, D is a covalent bond or a linking group linking Y to L or Z. The linking group may optionally be-O-, -S-, arylene, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl,carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulphonamido or amino.

R₄-R₇Each independently is hydrogen, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, azide, azo, phosphate, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido or amino.

Herein, the phrase "linking group" describes a group (e.g., a substituent) that is attached to two or more moieties in a compound.

Herein, the phrase "terminal group" describes a group (e.g., substituent) that is attached through one atom thereof to a single moiety in a compound.

When R is₄-R₆Each is hydrogen, and D is a covalent bond or a linking group, Y is an unsubstituted ethylene group attached (via D) to a pendant group described herein.

When R is₄-R₇Each is hydrogen (and D is R)₇) When Y is unsubstituted ethylene, not attached to a pendant group as described herein.

In some embodiments of any of the embodiments described herein, R is₄And R₅Each is hydrogen. These embodiments include polymeric backbones formed from a number of widely used vinyl monomers, including ethylene, including, for example, olefins (e.g., ethylene, propylene, 1-butene, isobutylene, 4-methyl-1-pentene), vinyl chloride, styrene, vinyl acetate, acrylonitrile, acrylates and derivatives thereof (e.g., acrylates, acrylamides), and methacrylates and derivatives thereof (e.g., methacrylates, methacrylamides).

As described hereinIn some embodiments of any of the embodiments of (1), R₆Is hydrogen. In some such embodiments, R₄And R₅Each is hydrogen.

In some embodiments of any of the embodiments described herein, R is₆Is methyl. In some such embodiments, R₄And R₅Each is hydrogen. In some such embodiments, the backbone units are units of a methacrylate or derivative thereof (e.g., methacrylate, methacrylamide).

In some embodiments of any of the embodiments described herein, the linking group represented by variable D is-O-, -C (= O) NH-, or phenylene. In exemplary embodiments, D is-C (= O) O-.

For example, when D is-O-, the backbone unit can optionally be a vinyl alcohol derivative (e.g., an ester or ether of a vinyl alcohol unit); when D is-C (= O) O-, an acrylate or methacrylate derivative (e.g. an ester of an acrylate or methacrylate unit); when D is-C (= O) NH-, acrylamide or methacrylamide units; and/or a styrene derivative (e.g. a substituted styrene unit) when D is phenylene.

In some embodiments of any of the embodiments described herein, L is a substituted or unsubstituted hydrocarbon from 1 to 10 carbon atoms in length. In some embodiments, the hydrocarbon is unsubstituted. In some embodiments, the hydrocarbon is a straight chain unsubstituted hydrocarbon, i.e., - (CH)₂)_i-, where i is an integer of 1 to 10.

In some embodiments of any of the embodiments described herein, L is substituted or unsubstituted ethylene. In some embodiments, L is unsubstituted ethylene (-CH)₂CH₂-)。

In some embodiments of any of the embodiments described herein, B is an oxygen atom. In some such embodiments, L is a hydrocarbon according to any of the respective embodiments described herein (i.e., L is not absent), and Z is a phosphate group attached to L.

In some embodiments of any of the embodiments described herein, B is absent. In some such embodiments, L is a hydrocarbon according to any of the respective embodiments described herein (i.e., L is not absent), and Z is a phosphonate group attached to L. In some embodiments, L is also absent such that the phosphorus atom of formula II is directly attached to Y.

In some embodiments of any of the embodiments described herein, a is a substituted or unsubstituted hydrocarbon from 1 to 4 carbon atoms in length.

In some embodiments of any of the embodiments described herein, a is an unsubstituted hydrocarbon. In some such embodiments, the unsubstituted hydrocarbon is 1 to 4 carbon atoms in length. In some embodiments, the hydrocarbon is a straight chain unsubstituted hydrocarbon, i.e., - (CH)₂)_j-, where j is an integer of 1 to 4.

In some embodiments of any of the embodiments described herein, a is substituted or unsubstituted ethylene.

In some of any of the embodiments described herein, a is unsubstituted ethylene (-CH)₂CH₂-). In such embodiments, the moiety having formula II (represented by variable Z) is similar to or the same as a phosphoethanolamine or phosphocholine moiety. Phosphoethanolamine and phosphocholine moieties are present in many naturally occurring compounds (e.g., phosphatidylcholine, phosphatidylethanolamine).

In some embodiments of any of the embodiments described herein, a is ethylene substituted with C-carboxy. In some embodiments, the C-carboxy group is attached to a carbon atom adjacent to the nitrogen atom shown in formula II (rather than to the oxygen atom shown). In this embodiment, the moiety having formula II (represented by variable Z) is similar or identical to a phosphoserine moiety. Phosphoserine is present in many naturally occurring compounds (e.g., phosphatidylserine).

Without being bound by any particular theory, it is believed that moieties similar or identical to naturally occurring moieties (e.g., phosphorylcholine, phosphoethanolamine, and/or phosphoserine) may be particularly biocompatible.

In some embodiments of any of the embodiments described herein, R is₁-R₃(substituents for nitrogen atom shown in formula II) are each independently hydrogen or C_1-4-an alkyl group. In some embodiments, R₁-R₃Each independently hydrogen or methyl. In some embodiments, R₁-R₃Each is methyl. In some such embodiments, R₁-R₃Each is hydrogen.

The variable n can be considered to represent the number of backbone units (represented by the variable Y) that are substituted with side groups represented by (-L-Z), and the variable m can be considered to represent the number of backbone units that are not substituted with such side groups. The sum n + m may be considered to represent the total number of backbone units in the polymeric backbone. The ratio n/(n + m) can be considered to represent the fraction of backbone units substituted by the side group represented by (-L-Z).

The backbone unit Y substituted with a pendant group can be the same as or different from the backbone unit Y that is not substituted with a pendant group (e.g., when m is at least 1).

The plurality of backbone units Y (represented by the variable n) substituted with a pendant group may be the same as or different from each other.

In addition, a plurality of pendant groups (-L-Z) attached to a plurality of backbone units Y (represented by the variable n) may be the same as or different from each other (e.g., A, B, R)₁、R₂、R₃And the identity of any one or more of L may be different).

In any of the embodiments described herein in which more than one backbone unit Y is not substituted with a pendant group described herein (i.e., when m is greater than 1), the plurality of backbone units Y (represented by the variable m) substituted with a pendant group can be the same as or different from each other.

The number of types of backbone units substituted with a pendant group, the number of types of backbone units not substituted with a pendant group (if any such units are present), and/or the number of types of pendant groups in the polymeric moiety can each independently be any number (e.g., 1,2, 3, 4, 5, 6, 7, 8, 9, 10, or greater).

In some embodiments of any of the embodiments described herein, the polymeric moiety is a copolymer moiety, i.e., the polymeric moiety comprises at least two different types of monomer units. The different types of monomeric units may differ in whether they comprise a pendant group (-L-Z) according to any of the respective embodiments described herein (e.g., when m is at least 1), in the type of backbone unit Y, and/or in the type of pendant group (-L-Z).

For example, in some of any of the embodiments described herein, the backbone units Y in each Y (-L-Z) unit can optionally be the same or different, while the L and Z moieties are the same in the Y (-L-Z) units. In some such embodiments, the backbone units not substituted by a pendant group (if any such units are present) may optionally be the same as backbone unit Y in each Y (-L-Z) unit. Alternatively, the backbone units not substituted by a pendant group (if any such units are present) may optionally be different from the backbone units Y in each Y (-L-Z) unit (and optionally the same in all backbone units not substituted by a pendant group).

In some of any of the embodiments described herein, the L moieties in each Y (-L-Z) unit may optionally be the same or different, while the backbone units Y and Z moieties are the same in the Y (-L-Z) unit. In some such embodiments, the backbone units not substituted by a pendant group (if any such units are present) may optionally be the same as backbone unit Y in each Y (-L-Z) unit. Alternatively, the backbone units not substituted by a pendant group (if any such units are present) may optionally be different from the backbone units Y in each Y (-L-Z) unit (and optionally the same in all backbone units not substituted by a pendant group).

In some of any of the embodiments described herein, the Z moiety in each Y (-L-Z) unit may optionally be the same or different, while the backbone units Y and Z moieties are the same in the Y (-L-Z) unit. In some such embodiments, the backbone units not substituted by a pendant group (if any such units are present) may optionally be the same as backbone unit Y in each Y (-L-Z) unit. Alternatively, the backbone units not substituted by a pendant group (if any such units are present) may optionally be different from the backbone units Y in each Y (-L-Z) unit (and optionally the same in all backbone units not substituted by a pendant group).

In any of the embodiments described herein in which the polymeric moiety is a copolymer moiety, any two or more different types of monomer units may be randomly or non-randomly distributed throughout the polymeric moiety. When the different types of monomer units are non-randomly distributed, the copolymer can be a copolymer characterized by any non-random distribution, such as an alternating copolymer, a periodic copolymer, and/or a block copolymer.

In some embodiments of any of the embodiments described herein, at least a portion of the monomeric units of the polymeric moiety comprise a targeting moiety (according to any of the embodiments described herein involving a targeting moiety).

The targeting moiety may optionally be comprised by (and optionally consist of) a backbone unit Y according to any respective embodiment described herein, a linking moiety L according to any respective embodiment described herein, and/or a moiety Z according to any respective embodiment described herein, e.g. wherein a substituent according to any respective embodiment described herein comprises a targeting moiety. For example, in which at least a portion of the backbone units Y have the formula-CR₄R₅-CR₆In some embodiments of D- (as described herein in any of the respective embodiments), R₄-R₆And D (optionally wherein D is R as described herein)₇) Comprising a targeting moiety according to any of the respective embodiments described herein (e.g., wherein R is₄-R₆And D is a substituted group comprising a substituent as a targeting moiety), and optionally any one or more R₄-R₆And D is a targeting moiety. However, many other structures comprising monomeric units containing substituents of (and optionally consisting of) targeting moieties are also encompassed by embodiments of the present invention.

When Y is a backbone unit not linked to L or Z (i.e., to a pendant group as described herein), D is R₇(end groups, as defined herein); and when Y is a master connected to L or ZIn the case of a chain unit, D is a covalent bond or a linking group linking Y to L or Z. The linking group can optionally be-O-, -S-, arylene, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido, or amino.

R₄-R₇Each independently is hydrogen, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, halo, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, sulfinyl, sulfonyl, cyano, nitro, azide, azo, phosphate, phosphonyl, phosphinyl, oxo, carbonyl, thiocarbonyl, urea, thiourea, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, C-carboxy, O-carboxy, sulfonamido or amino.

In some embodiments, the polymeric moiety is a copolymer moiety, wherein at least one type of monomeric unit comprises a targeting moiety (according to any of the respective embodiments described herein), and at least one type of monomeric unit does not comprise such a targeting moiety. The distribution of the monomer units comprising the targeting moiety can be according to any distribution described herein (e.g., random, alternating, periodic copolymers, and/or block copolymers) of the monomer units in the copolymer portion.

In some embodiments of any of the embodiments described herein wherein a portion of the monomeric units comprise a targeting moiety, the monomeric units comprising the targeting moiety are on average closer to the end of the polymeric moiety distal to the lipid moiety, e.g., the average distance (measured in atoms or backbone units along the backbone of the polymeric moiety) of the monomeric units comprising the targeting moiety from the lipid moiety is greater than the average distance of the other monomeric units from the lipid moiety.

In some embodiments, at least a portion (and optionally all) of the monomeric units comprising the targeting moiety form a block (of one or more monomeric units) near (and optionally at) the end of the polymeric moiety distal to the lipid moiety. In some such embodiments, the copolymer moiety contains a single monomeric unit comprising the targeting moiety, and the monomeric unit is terminal to the polymeric moiety distal to the lipid moiety.

Without being bound by any particular theory, it is hypothesized that a targeting moiety located distal to a lipid moiety may be more effective as a targeting moiety (e.g., bind to a target) e.g., because the targeting moiety is less spatially shielded (e.g., by a surface associated with the lipid moiety) and thus more exposed to and thus able to better contact a target in an aqueous environment.

In an alternative embodiment, the polymeric moiety does not comprise a targeting moiety as described herein according to any of the respective embodiments.

In some embodiments of any of the embodiments described herein, the percentage (represented by the formula 100% × n/(n + m)) of backbone units (represented by the variable Y) substituted with a pendant group represented by (-L-Z) is at least 20%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 30%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 40%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 50%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 60%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 70%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 80%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 90%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 95%. In some embodiments, the percentage of backbone units substituted with the foregoing pendant groups is at least 98%.

In some embodiments of any of the embodiments described herein, m is 0, such that each backbone unit (represented by the variable Y) is substituted with a pendant group represented by (-L-Z).

In some embodiments of any of the embodiments described herein, n is at least 5. In some embodiments, n is at least 10. In some embodiments, n is at least 15.

In some embodiments of any of the embodiments described herein, n is in the range of 2 to 1,000. In some embodiments of any of the embodiments described herein, n is in the range of 2 to 500. In some embodiments of any of the embodiments described herein, n is in the range of 2 to 200. In some embodiments of any of the embodiments described herein, n ranges from 2 to 100. In some embodiments of any of the embodiments described herein, n is in the range of 2 to 50. In some such embodiments, m is 0.

In some embodiments of any of the embodiments described herein, n is in the range of 3 to 1,000. In some embodiments of any of the embodiments described herein, n is in the range of 3 to 500. In some embodiments of any of the embodiments described herein, n is in the range of 3 to 200. In some embodiments of any of the embodiments described herein, n ranges from 3 to 100. In some embodiments of any of the embodiments described herein, n is in the range of 3 to 50. In some embodiments of any of the embodiments described herein, n is in the range of 5 to 50. In some embodiments of any of the embodiments described herein, n is in the range of 10 to 50. In some embodiments of any of the embodiments described herein, n is in the range of 10 to 25. In some such embodiments, m is 0.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 1,000. In some such embodiments, n is in the range of 2 to 1,000, such that the total number of backbone units is in the range of 2 to 2,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 500. In some such embodiments, n is in the range of 2 to 1,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 200. In some such embodiments, n is in the range of 2 to 1,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 100. In some such embodiments, n is in the range of 2 to 1,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 50. In some such embodiments, n is in the range of 2 to 1,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 20. In some such embodiments, n is in the range of 2 to 1,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

In some embodiments of any of the embodiments described herein, m is in the range of 0 to 10. In some such embodiments, n is in the range of 2 to 1,000. In some such embodiments, n is in the range of 3 to 1,000. In some embodiments, n is in the range of 3 to 500. In some embodiments, n is in the range of 3 to 200. In some embodiments, n is in the range of 3 to 100. In some embodiments, n is in the range of 3 to 50. In some embodiments, n is in the range of 5 to 50. In some embodiments, n is in the range of 10 to 50.

The lipid moiety according to any embodiment herein (represented by variable X in formula I herein) may be linked to the polymeric moiety according to any embodiment described herein in relation to the polymeric moiety.

The lipid moiety may optionally be derived from any lipid known in the art (including, but not limited to, naturally occurring lipids). The lipid moiety derived from the lipid may optionally consist of a hydrogen atom substituted at any position of the lipid with a polymeric moiety represented in formula I by [ Y (-L-Z) ] nY ] m (i.e., a polymeric moiety represented by formula Ia).

In some embodiments of any of the embodiments described herein, the lipid moiety (according to any of the respective embodiments described herein) is linked to a Y (-L-Z) unit (according to any of the embodiments described herein involving Y, L and/or Z), i.e., a backbone unit substituted with a pendant group described herein (e.g., rather than a backbone unit that is not substituted with a pendant group).

Alternatively or additionally, in some embodiments of any of the embodiments described herein (wherein m is at least 1), the lipid moiety (according to any of the respective embodiments described herein) may optionally be linked to a backbone unit (Y) that is not substituted with a pendant group described herein (e.g., instead of being linked to a backbone unit that is substituted with a pendant group). For example, the polymeric moiety may optionally be a copolymer, wherein the identity of the backbone unit attached to the lipid moiety varies randomly between molecules. Thus, the depiction of X in formula I attached to a backbone unit substituted with a pendant group (i.e., Y- (L-Z)) rather than to an unsubstituted backbone unit Y is arbitrary and not intended to be limiting.

In some embodiments of any of the embodiments described herein, the lipid moiety is a moiety of a lipid that is a fatty acid, a monoglyceride, a diglyceride, a triglyceride, a glycerophospholipid, a sphingolipid, or a sterol. In some embodiments, the lipid is a glycerophospholipid.

In some embodiments of any of the embodiments described herein, the lipid moiety comprises at least one fatty acid moiety (e.g., an acyl group derived from a fatty acid). The fatty acid moiety may be derived from saturated or unsaturated fatty acids. For example, the lipid moiety may consist of a fatty acid moiety, or a monoglyceride moiety comprising one fatty acid moiety, a diglyceride moiety comprising two fatty acid moieties, or a triglyceride moiety comprising three fatty acid moieties.

Examples of fatty acid moieties that may optionally be comprised by a lipid moiety include, but are not limited to, lauroyl, myristoyl, palmitoyl, stearoyl, palmitoyl, oleoyl, and linoleoyl.

Suitable examples of glycerophospholipids include, but are not limited to, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, and phosphatidylinositol.

In some embodiments of any of the embodiments described herein, the lipid moiety represented by the variable X has the general formula III:

formula III

W₁And W₂Each independently is hydrogen, alkyl, alkenyl, alkynyl or acyl, wherein W is₁And W₂Is not hydrogen;

j is-P (= O) (OH) -O-, or J is absent (such that K is directly connected to the indicated oxygen atom of the glycerol moiety);

k is a substituted or unsubstituted hydrocarbon of 1 to 10 carbon atoms in length; or K is absent (such that M is directly attached to J, or when J is absent, M is directly attached to the indicated oxygen atom of the glycerol moiety);

m is a linking group which is-O-, -S-, amino, sulfinyl, sulfonyl, phosphate, phosphonyl, phosphinyl, carbonyl, thiocarbonyl, urea, thiourea, carbamoyl, thiocarbamoyl, amido, carboxy or sulfonamide, or M is absent; and

q is a substituted or unsubstituted hydrocarbon from 1 to 10 carbon atoms in length and is attached to a backbone unit of the polymeric backbone according to any of the respective embodiments described herein.

When M is absent, K is also absent and Q is directly connected to J, or when J is absent, Q is directly connected to the depicted oxygen atom of the glycerol moiety.

In some of any of the embodiments described herein, W is₁And W₂One is hydrogen and the other is not hydrogen.

In some of any of the embodiments described herein, W is₁And W₂Are not hydrogen.

In some of any of the embodiments described herein, W is₁And W₂Is an alkyl, alkenyl, alkynyl or acyl group, which is 10 to 30 carbon atoms in length. In some embodiments, W₁And W₂Each of which is 10 to 30 carbon atoms in length.

Can optionally be used independently as W₁And/or W₂Examples of acyl groups of (a) include, but are not limited to, lauroyl, myristoyl, palmitoyl, stearoyl, palmitoyl, oleoyl, and linoleoyl.

In some embodiments of any of the embodiments described herein, J is-P (= O) (OH) -O- (e.g., the lipid moiety is a glycerophospholipid).

Herein, the length of the hydrocarbon represented by the variable K refers to the number of atoms separating J and M (i.e., along the shortest path between J and M), as shown in formula III.

When K is a substituted hydrocarbon, M may be attached to a carbon atom of the hydrocarbon itself, or to a substituent of the hydrocarbon.

In some of any of the embodiments described herein, K is an ethanolamine moiety (e.g., -CH)₂-CH₂-NH-, or-CH bound to a nitrogen atom₂-CH₂-, a serine moiety (e.g., -CH)₂-CH(CO₂H) -NH-, or-CH bound to a nitrogen atom₂-CH(CO₂H) -), glycerol moieties (e.g., -CH (OH) -CH-O-) and inositol moieties (e.g., -cyclohexyl (OH)₄-O-). In some embodiments, J is-P (= O) (OH) -O-.

In some embodiments of any of the embodiments described herein, M is amido, optionally-C (= O) NH-.

In some embodiments, the nitrogen atom of the amido group is attached to K. In some such embodiments, K is an ethanolamine or serine moiety as described herein.

In some embodiments of any of the embodiments described herein, J is absent (e.g., wherein the glycerolipid is not a glycerophospholipid). In some such embodiments, K is also absent such that M is directly attached to the depicted oxygen atom of the glycerol moiety, or Q is directly attached to the depicted oxygen atom of the glycerol moiety when M is also absent (e.g., where the glycerol lipid is a monoacylglycerol derivative or a diacylglycerol derivative). In some embodiments, M is a carbonyl linking group such that the attachment of M to the aforementioned oxygen atom of the glycerol moiety is through an ester linkage.

In some of any of the embodiments described herein, Q is substituted or unsubstituted methylene. In some such embodiments, M comprises a carbonyl (i.e., C (= O)) linking group. In some embodiments, M is an amido group (which contains a carbonyl group and a nitrogen atom). In some embodiments, C (= O) (e.g., C (= O of amido)) is linked to Q. In some embodiments, M consists of a carbonyl linking group.

In some embodiments of any of the embodiments described herein, Q is methylene substituted with one or two substituents. In some embodiments, the methylene group is substituted with one or two alkyl groups (e.g., C)_1-4-alkyl) substitution.

In some embodiments of any of the embodiments described herein, Q is methylene substituted with two substituents. In some embodiments, the methylene group is substituted with two alkyl groups (e.g., C)_1-4-alkyl) substitution. In some embodiments, alkyl is methyl such that Q is dimethylmethylene (-C (CH)₃)₂-)。

As exemplified in the examples section herein, substituted methylene groups (e.g., disubstituted methylene groups) represented by the variable Q are particularly suitable for participating in a polymerization reaction (e.g., as an initiator) because the radicals and/or ions on the methylene groups can be stabilized by one or more substituents thereof.

As further exemplified herein, the formation of an amido group (represented by the variable M) can serve as a convenient means of linking the substituted methylene group described above to a lipid (e.g., a naturally occurring lipid), such as phosphatidylethanolamine or phosphatidylserine.

As further exemplified herein, formation of an ester bond between a carbonyl group (e.g., comprised by M) and an oxygen atom of a lipid (e.g., an oxygen atom of a glycerol moiety) can serve as a convenient means of attaching the substituted methylene group described above to a lipid (e.g., a naturally occurring lipid), such as a monoacylglycerol, diacylglycerol, phosphatidylglycerol, or phosphatidylinositol.

A targeting moiety:

as noted above, in some embodiments of any of the embodiments described herein, at least a portion of the monomeric units comprise a targeting moiety (according to any of the embodiments described herein involving a targeting moiety).

Herein, a "targeting moiety" refers to a moiety that is capable of bringing a compound (e.g., a compound according to some embodiments of the present invention) into proximity with a selected substance and/or material (referred to herein as a "target"). The target is optionally a cell (e.g., a proliferative cell associated with a proliferative disease or disorder), wherein access is such that the targeting moiety facilitates attachment and/or internalization of the compound into the target cell and such that the compound can exert a therapeutic effect.

In any of the embodiments described herein, wherein m is at least 1, according to any of the respective embodiments described herein, at least a portion of the monomer units comprising the targeting moiety (the number of which is represented by the variable m) are monomer units that do not comprise a pendant group represented by (-L-Z). In some such embodiments, each monomer unit comprising a targeting moiety (according to any of the respective embodiments described herein) is a monomer unit comprising a pendant group represented by (-L-Z) (i.e., a backbone unit Y substituted with (-L-Z)), i.e., none of the monomer units comprising a pendant group represented by (-L-Z) comprises a targeting moiety as described above.

In any of the embodiments described herein wherein m is at least 1, each monomer unit (the number of which is represented by the variable m) that does not comprise a pendant group represented by (-L-Z) comprises a targeting moiety (according to any of the respective embodiments described herein). In some such embodiments, each monomer unit comprising a targeting moiety (according to any of the respective embodiments described herein) is a monomer unit that does not comprise a pendant group represented by (-L-Z), i.e., none of the monomer units comprising a pendant group represented by (-L-Z) comprises the above-described targeting moiety, and each monomer unit that does not comprise a pendant group represented by (-L-Z) comprises the above-described targeting moiety.

In any of the embodiments described herein wherein m is at least 1, the monomer unit comprising the targeting moiety may consist essentially of the backbone unit Y (according to any of the respective embodiments described herein) substituted with one or more targeting moieties (according to any of the respective embodiments described herein).

The backbone unit Y of the monomeric unit comprising the targeting moiety may optionally be structurally different (optionally significantly different) from the backbone unit Y of the other monomeric units in the polymeric moiety (according to any of the respective embodiments described herein).

In any of the embodiments described herein wherein m is at least 1, the polymeric moiety comprises a monomeric unit comprising a targeting moiety, and the monomeric unit is terminal to the polymeric moiety distal to the lipid moiety. In such embodiments, the compound represented by formula I has formula Ib:

formula Ib

Wherein:

t is a monomeric unit comprising a targeting moiety (according to any of the respective embodiments described herein);

x and T are linked to the distal end of the moiety represented by [ Y (-L-Z) ] nY m-1; and

x, Y, L, Z, n and m are defined according to any of the embodiments described herein with reference to formula I, provided that m is at least 1.

It is to be understood that T in formula Ib is the type of monomer unit represented by Y in formulas I and Ia (i.e., does not have a pendant group represented by (-L-Z)), and that the number of monomer units represented by Y rather than T (i.e., does not have a pendant group represented by (-L-Z)) is represented by the value m-1, such that the total number of monomer units that do not have a pendant group represented by (-L-Z), including T, as in formulas I and la, is represented by the variable m.

In some embodiments, m is 1 such that m-1 is 0, and the compound represented by formula Ib thus has the formula: x- [ Y (-L-Z) ] n-T, wherein L, T, X, Y, Z and n are defined according to any embodiment described herein.

A monomeric unit comprising a targeting moiety according to any of the respective embodiments described herein may optionally be prepared as follows: the method may comprise preparing a monomer comprising the targeting moiety and using the monomer to prepare a polymeric moiety as described herein (e.g., by polymerization of a monomer according to any of the respective embodiments described herein) and/or modifying a monomer unit in a polymeric moiety after preparing a polymeric moiety (e.g., by polymerization of a monomer according to any of the respective embodiments described herein) using any suitable technique known in the art, including but not limited to conjugation techniques.

In some embodiments of any of the embodiments described herein relating to a targeting moiety, the targeting moiety does not comprise a moiety having general formula II (according to any of the respective embodiments described herein). For example, even if the moiety represented by formula II is capable of forming a bond with a target as described herein, in some embodiments, the phrase "targeting moiety" should be understood to refer to a moiety that is different from the moiety represented by the variable Z (having formula II).

In some embodiments of any one of the embodiments described herein, the pendant group represented by (-L-Z) is selected such that no bond is formed with the target and/or such that the structure and/or properties of the targeting moiety described herein in any one of the respective embodiments are not included. For example, in embodiments in which a targeting moiety comprising a nucleophilic group (according to any of the respective embodiments described herein), e.g., an amine group, is capable of forming a bond (e.g., covalent bond) with a target, the variable Z (having formula II) is optionally selected such that the described amine/ammonium group is a tertiary amine/ammonium (i.e., R₁-R₃No more than one of which is hydrogen) or quaternary ammonium (i.e., R)₁-R₃Neither is hydrogen), preferably quaternary ammonium (e.g., containing trimethyl amino groups, such as in phosphorylcholine). Tertiary amine groups, and particularly quaternary ammonium groups, may be nucleophilic groups that are significantly less reactive than primary and secondary amine groups.

In some embodiments of any of the embodiments described herein that relate to a targeting moiety, the targeting moiety comprises (and optionally consists of) at least one functional group capable of forming a covalent bond or a non-covalent bond (preferably a selective non-covalent bond) with a substance and/or material (referred to herein as "target"), e.g., at the surface of the target (e.g., the surface of a cell and/or tissue).

Herein, the phrase "functional group" includes chemical groups and moieties of any size and any functionality described herein (e.g., any functionality capable of forming a covalent or non-covalent bond with a target).

Non-covalent bonds according to any of the respective embodiments described herein may optionally be achieved by non-covalent interactions, such as, but not limited to, electrostatic attraction, hydrophobic bonds, hydrogen bonds, and aromatic interactions.

In some embodiments, the targeting moiety comprises a functional group capable of forming a non-covalent bond selective for the target, e.g., the affinity of the targeting moiety and/or functional group for the target (e.g., as determined based on the dissociation constant) is greater than the affinity of the targeting moiety and/or functional group for most (or all) other compounds capable of forming a non-covalent bond with the targeting moiety.

In some embodiments of any of the embodiments described herein, the one or more functional groups are capable of forming a covalent bond with one or more specific functional groups (e.g., hydroxyl, amine, thiol, and/or oxo groups) present on a target (e.g., a target according to any of the respective embodiments described herein).

Examples of functional groups (in the targeting moiety) capable of forming a covalent bond with a target (according to any of the respective embodiments described herein) and the types of covalent bonds they are capable of forming include, but are not limited to:

nucleophilic groups such as thiol, amine (e.g., primary or secondary amine), and hydroxyl groups, which can form covalent bonds with, for example, nucleophilic leaving groups (e.g., any nucleophilic group described herein), michael acceptors (e.g., any michael acceptors described herein), acid halides, isocyanates, and/or isothiocyanates (e.g., as described herein) in the target;

nucleophilic leaving groups such as halogens, azides (-N)₃) Sulfates, phosphates, sulfonyl groups (e.g., methanesulfonyl, toluenesulfonyl), N-hydroxysuccinimide (NHS) (e.g., NHS esters), sulfo-N-hydroxysuccinimide, and anhydrides, which can form covalent bonds with, for example, nucleophilic groups in a target (e.g., as described herein);

michael acceptors such as enones (e.g., maleimides, acrylates, methacrylates, acrylamides, methacrylamides), nitro and vinyl sulfones, which can form covalent bonds with, for example, nucleophilic groups (e.g., as described herein), optionally thiol groups, in a target;

a dihydroxyphenyl group (according to any of the respective embodiments described herein) that can form a covalent bond with, for example, a nucleophilic group (e.g., as described herein) and/or a substituted or unsubstituted phenyl group (e.g., another dihydroxyphenyl group) in a target as described herein;

an acyl halide (-C (= O) -halogen), isocyanate (-NCO), and isothiocyanate (-N = C = S) group, which can form a covalent bond with, for example, a nucleophilic group in a target (e.g., as described herein);

a carboxylic acid (-C (= O) OH) group which can form a covalent bond with, for example, a hydroxyl group in the target to form an ester bond and/or an amine group (e.g. a primary amine) in the target to form an amide bond (optionally by reaction with a coupling reagent such as a carbodiimide); and/or the carboxylic acid group is in the target and can form an amide or ester bond with an amine or hydroxyl group in the targeting moiety, respectively;

an oxo group (optionally in an aldehyde group (-C (= O) H)) that can form a covalent imine bond with an amine group (e.g., a primary amine) in the target; and/or oxo groups (optionally in aldehyde groups) in the target and may form covalent imine bonds with amine groups in the targeting moiety; and/or

A thiol group which can form a covalent disulfide (-S-S-) bond with the thiol group in the target.

Modification of the monomeric (e.g., prior to polymerization) or monomeric units of the polymeric moiety (e.g., after polymerization) to include any of the functional groups described herein can optionally be performed using any suitable conjugation technique known in the art. The skilled person will be able to readily select a suitable technique for any given molecule to be modified.

As used herein, the term "dihydroxyphenyl" refers to an aryl group (as defined herein) which is a phenyl group substituted at any position thereof with two hydroxy groups. The phenyl group may be optionally substituted with additional substituents (which may optionally comprise additional hydroxyl groups) to form a substituted dihydroxyphenyl group; or alternatively, the phenyl group contains no substituents other than the two hydroxyl groups, such that the dihydroxyphenyl group is an unsubstituted dihydroxyphenyl group.

In some embodiments of any one of the embodiments described herein, the dihydroxyphenyl group is an ortho-dihydroxyphenyl group (in which the hydroxyl group is attached to the phenyl group at adjacent positions) or a para-dihydroxyphenyl group (in which the hydroxyl group is attached to the opposite side of the phenyl ring), each being a substituted or unsubstituted dihydroxyphenyl group. In some such embodiments, the o-dihydroxyphenyl group or the p-dihydroxyphenyl group is an unsubstituted dihydroxyphenyl group.

According to Lee et alPNAS 2006, 103:12999-13003]Brodie, etc. [Biomedical Materials2011, 6:015014]And/or international patent application PCT/IL2015/050606 (published as WO2015/193888) for any one or more of the attachment mechanisms described for the dihydroxyphenyl (catechol) group, the dihydroxyphenyl groups according to any of the respective embodiments described herein may optionally be covalently and/or non-covalently bonded to a target, the contents of each of which are incorporated in their entirety, and in particular with respect to the bond formed by the dihydroxyphenyl (catechol) group with a surface.

In some embodiments of any of the embodiments described herein, the functional group capable of forming a bond with the target is a functional group capable of forming a covalent bond with an amine group, optionally a primary amine group. In some such embodiments, the target comprises one or more amino acids or amino acid residues, such as a peptide or polypeptide of any length (e.g., at least two amino acid residues, such as a protein), and the amine group can optionally be a lysine side chain amine group and/or an N-terminal amine group. In some embodiments, the target comprises an extracellular matrix protein, such as collagen. In some embodiments, the target comprises cartilage (e.g., articular cartilage).

In some embodiments of any one of the embodiments described herein, the targeting moiety comprises (and optionally consists of) at least one functional group capable of forming a non-covalent bond with the target (e.g., as described herein in any one of the respective embodiments).

In some embodiments of any one of the embodiments described herein, the functional group capable of forming a non-covalent bond with a target comprises (and optionally consists of) a polysaccharide and/or a polypeptide (e.g., a protein and/or a fragment thereof), wherein the target optionally comprises a ligand for the polysaccharide and/or polypeptide; and/or the target comprises a polysaccharide and/or polypeptide (e.g., a protein and/or fragment thereof), and the functional group capable of forming a non-covalent bond with the target is a ligand for the polysaccharide and/or polypeptide.

Examples of suitable polysaccharides and/or polypeptides and their ligands include, but are not limited to:

avidin or streptavidin as the polypeptide described herein, and biotin as its ligand;

a polysaccharide-binding polypeptide as the polypeptide described therein, and a complementary polysaccharide as its ligand (or a complementary polysaccharide-binding polypeptide as a ligand for a polysaccharide described herein);

a collagen-binding polypeptide that is a polypeptide described therein, and a complementary collagen that is a ligand thereof (or a collagen that is a polypeptide described herein and a complementary collagen-binding polypeptide that is a ligand thereof);

a cellular receptor expressed by the cell and a ligand selectively bound by the receptor;

an antibody to any antigen (e.g., wherein the target described herein optionally comprises the antigen) as a polypeptide as described herein or a fragment of such an antibody, and the corresponding antigen as its ligand; and

an antibody mimetic of any antigen (e.g., wherein the target described herein optionally comprises the antigen).

Examples of cellular receptors expressed by cells include, but are not limited to, receptors characteristic of particular types of cells and/or tissues, and receptors overexpressed by cancer cells. The cellular receptor or cell is optionally a target as described herein, and the targeting moiety optionally comprises any ligand for the receptor. Examples of such ligands include, but are not limited to, transferrin; a ligand for transferrin receptor, which can optionally target transferrin receptor overexpressed by some cancer cells; keratinocyte growth factor (KGF or FGF7), which is specific for cells of epithelial origin and which may optionally target KGF receptors, such as KGF receptors overexpressed by endometrial or pancreatic cancer [ Visco et al,Int J Oncol 1999, 15:431-435; Siegfried et al., Cancer 1997, 79:1166-1171](ii) a And Epidermal Growth Factor (EGF), which may optionally target an EGF receptor, optionally erbB, e.g., which is overexpressed by gliomas and endometrial cancers [ Normanno et al,Curr Drug Targets2005, 6:243-257])。

as used herein, the term "antibody" includes any type of immunoglobulin.

As used herein, the phrase "antibody mimetic" includes any type of molecule, optionally a polypeptide, referred to in the art as capable of selectively binding (e.g., non-covalently) an antigen. Non-limiting examples of antibody mimetics include affibody, affilin, affimer, affitin, alphabody, anticalin, avimer, DARPin, Fynomer, Kunitz domain peptide, and monobody, e.g., such as Nygren @FEBS J 2008, 275:2668-2676], Ebersbach et al. [J Mol Biol 2007, 372:172-185], Johnson et al. [Anal Chem 2012, 84:6553-6560], Krehenbrink et al. [J Mol Biol 2008, 383:1058-1068], Desmet et al. [Nature Comm 2014, 5:5237], Skerra [FEBS J 2008, 275:2677-2683], Silverman et al. [Nature Biotechnol 2005, 23:1556-1561], Stumpp et al. [Drug Discov Today2008, 13:695-701], Grabulovski et al. [J Biol Chem 2007, 282:3196-3204], Nixon& Wood [Curr Opin Drug Discov Devel 2006, 9:261-268], Koide & Koide [Methods Mol Biol 2007, 325:95-109]And Gebauer& Skerra [Curr Opin Chem Biol2009, 13:245-255]The contents of each of which are incorporated in their entirety, and particularly with respect to a particular type of antibody mimetic.

As used herein, the phrase "polysaccharide binding polypeptide" includes any polypeptide or oligopeptide (peptide chain of at least 2, and preferably at least 4 amino acid residues in length) that is capable of selectively binding (e.g., non-covalently) to a polysaccharide. Various polysaccharide binding polypeptides and their binding specificities are known to those skilled in the art and include short peptide sequences (e.g., 4 to 50, optionally 4 to 20 amino acid residues in length) and longer polypeptides such as proteins or fragments thereof (e.g., carbohydrate binding modules and/or domains). In addition, the phrase "polysaccharide binding polypeptide" includes antibodies that are capable of specifically binding to a polysaccharide. Such antibodies are available to the skilled person and/or the skilled person will know how to prepare such antibodies using immunological techniques known in the art.

Examples of polysaccharide-binding polypeptides that may be used in some of any of the embodiments of the present invention include, but are not limited to, carbohydrate-binding modules (CBMs); and hyaluronic acid binding peptides, polypeptides and/or modules (e.g., having a structure as described in International patent application publication WO 2013/1156; International patent application publication WO 2014/071132; Barta et alBiochem J 1993, 292:947-949], Kohda et al. [Cell 1996, 86:767-775], Brisset & Perkins [FEBS Lett 1996, 388:211-216], Peach et al. [J Cell Biol 1993, 122:257-264], Singh et al. [Nature Materials 2014, 13:988-995]And Zaleski et alAntimicrob Agents Chemother 2006, 50:3856-3860]The sequence of any one of, wherein the contents of each are incorporated in their entirety, and in particular with respect to the contents of a particular polysaccharide binding polypeptide), e.g., GAHWQFNALTVR (SEQ ID NO: 1) (hyaluronic acid binding peptide sequence).

Examples of CBMs that may be used in some of any of the embodiments of the present invention include, but are not limited to, CBMs belonging to the following families: CBM3, CBM4, CBM9, CBM10, CBM17 and/or CBM28 (which may optionally be used to bind cellulose, for example, in a cellulose-containing target); CBM5, CBM12, CBM14, CBM18, CBM19 and/or CBM33 (which may optionally be used to bind chitin and/or other polysaccharides comprising N-acetylglucosamine, for example in a chitin-containing target); CBM15 (optionally useful for binding hemicellulose, e.g., in hemicellulose-containing targets); and/or CBM20, CBM21, and/or CBM48 (which may optionally be used to bind starch and/or glycogen, e.g., in a starch-and/or glycogen-containing target).

As used herein, the phrase "collagen-binding polypeptide" includes any polypeptide or oligopeptide (peptide chain of at least 2, and preferably at least 4 amino acid residues in length) that is capable of selectively binding (e.g., non-covalently) to collagen (e.g., one type of collagen, some types of collagen, all types of collagen), including glycosylated polypeptides and oligopeptides, such as peptidoglycans and proteoglycans. Various collagen-binding polypeptides and their binding specificities will be known to the skilled person and include short peptide sequences (e.g. 4 to 50, optionally 4 to 20 amino acid residues in length) and longer polypeptides such as proteins or fragments thereof (e.g. collagen-binding domains). In addition, the phrase "collagen-binding polypeptide" includes antibodies that are capable of specifically binding to collagen. Such antibodies will be available to the skilled person and/or the skilled person will know how to prepare such antibodies using immunological techniques known in the art.

Examples of collagen-binding polypeptides that can be used in embodiments of the invention include, but are not limited to, collagen-binding proteins (e.g., decorin), fragments thereof, and/or collagen-binding polypeptides such as those described in U.S. Pat. No. 8,440,618, Abd-Elgaliel& Tung [Biopolymers 2013, 100:167-173], Paderi et al. [Tissue Eng Part A 2009, 15:2991-2999], Rothenfluh et al. [Nat Mater 2008, 7:248-254]And Helms et alJ Am Chem Soc 2009, 131:11683-11685](the contents of each of which are incorporated in their entirety, and in particular with respect to the contents of a particular collagen-binding polypeptide), e.g., the sequence WYRRGRL (SEQ ID NO: 2).

It is expected that during the life of a patent maturing from this application many relevant binding functionalities and moieties will be developed and/or revealed, and the scope of the terms "targeting moiety", "functional group", "cell receptor", "antibody mimetic", "collagen binding polypeptide" and "polysaccharide binding polypeptide" and the like is intended to include all such a priori new technologies.

In some embodiments of any of the embodiments described herein, the functional group in the targeting moiety (according to any of the respective embodiments described herein) is attached to a linking group (as defined herein). The linking group can optionally be any linking group or linking moiety described herein, including but not limited to substituted or unsubstituted hydrocarbons. In some embodiments, the targeting moiety (optionally a substituent of backbone unit Y) consists essentially of a functional group linked to the remainder of the polymeric moiety through a linking group.

The functional group can optionally be attached to the linking moiety through a covalent bond that can result from a reaction between two functional groups (e.g., any of the covalent bonds and/or functional groups described herein where a covalent bond is formed between the functional group and the target).

In some embodiments of any of the embodiments described herein that involve a functional group of a peptide or polypeptide, the amino acid residue of the peptide or polypeptide is optionally linked to a linking group of a targeting moiety, e.g., by an amide bond formed from an amine or carboxylic acid group in the peptide or polypeptide (e.g., in the N-terminus, lysine side chain, C-terminus, glutamic acid side chain and/or aspartic acid side chain), an ester bond formed from a hydroxyl or carboxylic acid group in the peptide or polypeptide (e.g., in the serine side chain, threonine side chain, C-terminus, glutamic acid side chain and/or aspartic acid side chain), and/or a disulfide bond formed from a thiol group in the peptide or polypeptide (e.g., in the cysteine side chain). In some embodiments, the amino acid residue attached to the linking group is an N-terminal and/or C-terminal residue, e.g., any amino acid residue attached through an N-terminal amino group or a C-terminal carboxylic acid group, and/or a terminal lysine, glutamic acid, aspartic acid, serine, threonine, and/or cysteine residue attached through a side chain thereof.

In some embodiments, amino acid residues and/or peptides (e.g., 2 to 20 amino acid residues in length) are added to the N-terminus and/or C-terminus of a functional group peptide or polypeptide sequence (according to any of the respective embodiments described herein), and the sequence is linked to a linking group. Examples of such terminal amino acid residues and/or peptides include, but are not limited to, glycine residues and peptides having terminal glycine residues, which can be used to attach a linking group to the N-terminus or C-terminus (according to any of the respective embodiments described herein); serine and threonine residues and peptides having terminal serine or threonine residues that can be used to attach a linking group to a hydroxyl group in a serine or threonine side chain, optionally through an ester linkage (according to any of the respective embodiments described herein); and cysteine residues and peptides having terminal cysteine residues, which can be used to attach a linking group to a peptide via a disulfide bond (according to any of the respective embodiments described herein).

In some embodiments, the peptide or polypeptide is linked to the linking group through a terminal amino acid residue to minimize interference (e.g., steric interference) with the functionality of the peptide or polypeptide upon linking to the linking group.

In some embodiments, linking a peptide or polypeptide to a linking group through a terminal glycine facilitates linking by minimizing interference (e.g., steric interference) of the amino acid side chain (glycine deficiency) with linking to the linking group.

Lipids and liposomes:

as described herein, a liposome according to embodiments comprises at least one bilayer-forming lipid.

Herein, the term "bilayer-forming lipid" includes any compound in which a bilayer may be formed from a purely aqueous solution of the compound, the bilayer comprising two parallel molecular layers of the compound (referred to as "lipid").

Typically, the bilayer (e.g. in liposomes according to some of any of the embodiments described herein) comprises relatively polar lipid moieties at both surfaces of the bilayer, which may optionally comprise an interface with an aqueous solution and/or an interface with a solid surface; and a relatively hydrophobic portion of the lipid inside the bilayer at the interface between the two lipid molecules forming the bilayer.

Examples of bilayer-forming lipids include glycerophospholipids (e.g., glycerophospholipids according to any of the respective embodiments described herein). It will be appreciated that the polymeric compound described herein may optionally be a bilayer-forming lipid, which may form a bilayer by itself or in combination with one or more additional bilayer-forming lipids.

In some embodiments of any one of the embodiments described herein, the bilayer-forming lipid comprises at least one charged group (e.g., one or more negatively charged groups and/or one or more positively charged groups).

In some embodiments, the bilayer-forming lipid is zwitterionic; including both (e.g., the same number of) negatively and positively charged groups (e.g., one each).

In some embodiments of any of the embodiments described herein, the molar ratio of bilayer-forming lipids (bilayer-forming lipids included in addition to the polymeric compound) and polymeric compound in the liposome is in the range of 5:1 to 5,000:1 (bilayer-forming lipid: polymeric compound), optionally in the range of 10:1 to 2,500:1, optionally in the range of 25:1 to 1,000:1, and optionally in the range of 50:1 to 500: 1.

In some embodiments of any of the embodiments described herein, the polymeric moiety in the liposome comprises a lipid moiety represented by variable X in formula I (according to any of the respective embodiments described herein), the lipid moiety comprising residues of a bilayer-forming lipid (e.g., a glycerophospholipid) comprised by or closely related to the bilayer-forming lipid comprised by the liposome in addition to the polymeric moiety, e.g., wherein the lipid moiety comprises glycerophospholipid residues and the liposome comprises another glycerophospholipid as the bilayer-forming lipid (e.g., optionally wherein the fatty acid residues in the glycerophospholipid residues are about the same length as the fatty acid residues in the bilayer-forming lipid, and optionally wherein the fatty acid residues in the glycerophospholipid residues are substantially the same as the fatty acid residues in the bilayer-forming lipid).

Without being bound by any particular theory, it is believed that the similarity between the lipid portion of the polymeric moiety and the bilayer-forming lipids helps to anchor the lipid portion of the polymeric moiety in the liposome comprising the bilayer-forming lipids.

Liposomes can optionally comprise a single bilayer (e.g., a unilamellar vesicle) or a plurality of bilayers (e.g., a multilamellar vesicle), wherein each bilayer optionally independently forms a closed vesicle, comprising, for example, concentric bilamellar vesicles and/or a plurality of individual bilamellar vesicles surrounded by the same bilamellar vesicle.

Liposomes according to any of the respective embodiments described herein can be approximately spherical, or can have any alternative shape, such as an elongated tube and/or a flat (e.g., sheet) shape.

The average diameter of the liposomes may optionally be in the range of 20nm to 1000nm, optionally in the range of 50nm to 300 nm.

In some embodiments of any of the embodiments described herein, the liposomes have an average diameter of at least 100nm, for example 100 to 1000nm, or 100nm to 300 nm. In some embodiments, the liposomes have an average diameter of at least 125nm, for example 125 to 1000nm, or 125nm to 300 nm. In some embodiments, the liposomes have an average diameter of at least 150nm, such as from 150 to 1000nm, or from 150nm to 300 nm. In some embodiments, the liposomes have an average diameter of at least 160nm, such as 160 to 1000nm, or 160nm to 300 nm. In some embodiments, the liposomes have an average diameter of at least 170nm, such as 170 to 1000nm, or 170nm to 300 nm.

As illustrated herein, liposome diameters of at least about 100nm (e.g., about 160-170nm) are associated with greater in vivo retention times.

The average diameter of the liposomes can be determined, for example, by dynamic light scattering measurements according to procedures known in the art (e.g., as described in the examples section herein).

In some embodiments of any of the embodiments described herein involving liposomes, the liposomes further comprise at least one functional moiety or agent (in addition to the therapeutically active agents described herein) incorporated into the liposome surface and/or within the lipid bilayer and/or core of the liposome (e.g., within and/or encapsulated by the liposome bilayer).

Examples of functional moieties and agents suitable for inclusion in the embodiments described herein include, but are not limited to, labeling moieties or agents, and/or targeting moieties or targeting agents (e.g., targeting moieties or agents on the surface of liposomes).

Examples of labeling moieties or agents include chromophoric (e.g., absorbing visible light), fluorescent, phosphorescent, and/or radioactive moieties and compounds. Many such compounds and moieties (and techniques for making such moieties) are known to the skilled artisan.

The targeting moiety in the liposome according to any of the respective embodiments described herein may optionally be a targeting moiety according to any of the respective embodiments described herein. The targeting moiety in the liposome may be comprised by a polymeric compound according to some embodiments of the invention (according to any of the respective embodiments described herein), said liposome comprising the polymeric compound. Alternatively or additionally, the targeting moiety in the liposome may optionally comprise another compound in the liposome, optionally a bilayer-forming lipid conjugated to a targeting moiety according to any of the respective embodiments described herein (according to any of the respective embodiments described herein).

Herein, a "targeting agent" refers to a compound ("agent") comprising (and optionally consisting essentially of) a targeting moiety according to any of the respective embodiments described herein (e.g., in the context of a targeting moiety comprised by a polymeric compound described herein). Generally, the phrase "targeting agent" is used to refer to a compound other than a polymeric compound comprising a targeting moiety as described herein.

In some embodiments, the functional moiety (e.g., targeting moiety or labeling moiety) is covalently attached to the liposome. In some embodiments, such attachment can be achieved by using techniques known in the art (e.g., amide bond formation).

Therapeutically active agents and indications:

as used herein, the phrase "therapeutically active agent" refers to any agent (e.g., compound) that has a therapeutic effect, as well as any agent moiety (e.g., compound moiety) that when released (e.g., upon cleavage of one or more covalent bonds) produces an agent with a therapeutic effect.

The therapeutically active agent added to the liposome and/or on the surface of the liposome can be, for example, attached to the liposome via covalent or non-covalent (e.g., electrostatic and/or hydrophobic) bonds (e.g., to the outer and/or inner surface of the liposome membrane), incorporated within the liposome membrane (e.g., lipophilic agents stably distributed to the lipid phase of the liposome), and/or encapsulated within the core of the liposome (e.g., hydrophilic agents in the aqueous compartment of the liposome).

In some embodiments, the therapeutically active agent is a moiety covalently attached to the liposome (e.g., attached to a lipid to form a lipid derivative comprising the moiety). In some embodiments, such attachment can be achieved by using techniques known in the art (e.g., amide bond formation).

In some embodiments of any of the embodiments described herein, the therapeutically active agent is, for example, an analgesic, an anti-inflammatory agent, an anti-proliferative agent, an antimicrobial agent (including an antibacterial agent, an anti-mycobacterial agent, an antiviral agent, an antifungal agent, an antiprotozoal agent, and/or an antiparasitic agent), and/or a vaccine antigen. In some such embodiments, the therapeutically active agent is an analgesic and/or an anti-inflammatory agent. In some such embodiments, the therapeutically active agent may be used alone or in combination with another therapeutically active agent for the treatment of osteoarthritis.

In some of any of the embodiments described herein, the liposomes of the embodiments of the invention are administered to a subject in need thereof with an additional therapeutically active agent or pharmaceutical composition comprising the same that is useful for treating a specified medical condition. Additional therapeutically active agents may be incorporated into the liposomes of the present embodiments, may be incorporated into other liposomes which may have the same or different agent retention times, or may simply be mixed with a suitable liposome-free carrier.

Examples of suitable analgesics include, but are not limited to, allylamine, α -methylfentanyl, AP-237, bezilimide, butorphanol, buprenorphine, carfentanil, clonidine, codeine, desmethylputamine, dextromoramide, dexocine, diphenoxylate, dihydrocodeine, dihydrotolorphine, dihydromorphine, diphenoxylate, dipivoperidone, esmoldoline, ethylmorphine, etorphine, fentanyl, isocodeine, hydrocodone, hydromorphone, ketamine, katomidone, lifloxamine, levomethadone (levomethadone), levomethamphetamine, levorphanol, loperamide, meptazinol, methadone, mexiletine, cephamyline, morphine, nalbuphine, hydroxyfentanil, oxycodone, oxymorphone, p-acetaminophenol, pentazocine, pethidine, phenethylphenylacetoxypiperidine, procarbazine, meperidine, propoxyphene, remifentanil, sufentanil, tapentadol, telidine and tramadol.

Non-steroidal anti-inflammatory agents (e.g., non-steroidal anti-inflammatory agents as described herein) as well as steroidal anti-inflammatory agents can also be used as analgesics.

Examples of suitable anti-inflammatory agents include, but are not limited to, alclofenac; alclometasone (e.g., alclometasone dipropionate); algestrol (e.g., pregnenide); an alpha amylase; an acerola; anxi nonspecific; amfenac (e.g., sodium amfenac); aminoprolise (e.g., aminoprolise hydrochloride); anakinra; anisic acid; anizanifen; azapropazone; aspirin; balsalazide disodium; benzyda; benoxaprofen; benzydamine (e.g., benzydamine hydrochloride); bromelain; bromopimox; budesonide; carprofen; fluorene propionic acid; cipentazone; chlorothioxuric acid; clobetasol (e.g., clobetasol propionate, clobetasol butyrate); clopidogrel acid; chlorthiokatron (chlorthiokatron propionate); cormethasone (acetic acid cormethasone); trutodo pine; deflazacort; (ii) donepezil; desoximetasone; dexamethasone (e.g., dexamethasone dipropionate); diclofenac (e.g., diclofenac potassium, diclofenac sodium); diflorasone (e.g., diflorasone diacetate); diflufenican (e.g., sodium diflufenican); diflunisal; difluprednate; biphthalic ketones; (ii) ciclesonide; emthylosol; (ii) an enromamab; enoxicam (e.g., enoxicam sodium); epiprazole; etodolac; etofenamate; felbinac; (ii) fenomod; fenbufen; fenchloric acid; benzoic acid; fendulx; fenpipalon; fentiacid; frazadone; fluzacort; flufenamic acid; fluorolumidazole; flunisolide (e.g., flunisolide acetate); flunixin (e.g., flunixin meglumine); fluorocyclobutane (e.g., fluorocarbutyl ester); fluorometholone (e.g., fluorometholone acetate); fluoroquinazone; flurbiprofen; fluretofen; fluticasone (e.g., fluticasone propionate); furaprofen; (ii) furobufen; halcinonide; halobetasol (e.g., halobetasol propionate); haloprednisone (e.g., haloprednisone acetate); isobutylphenylacetic acid; ibuprofen (e.g., aluminum ibuprofen, picoprofen); ilodap; indomethacin (e.g., indomethacin sodium); indoprofen; indoclovir; indazole; isoflupredone (e.g., isoflupredone acetate); isoxemic acid; isoxicam; ketoprofen; lofeimidazole (e.g., lofeimidazole hydrochloride); lornoxicam; loteprednol etabonate (e.g., loteprednol etabonate); meclofenamic acids (e.g., sodium meclofenamate, meclofenamic acid); (ii) meclosone (e.g., meclosone dibutyrate); mefenamic acid; mesalazine; mexilazone; methylprednisolone (e.g., methylprednisolone sulfoheptide); momifluorate; nabumetone; naproxen (e.g., naproxen sodium); naproxol; nimazone; olsalazine (e.g., olsalazine sodium); an superoxide dismutase; olpanoxin; oxaprozin; oxyphenbutazone; renitolin (e.g., renitolin hydrochloride); pentosan polysulfate salts (e.g., sodium pentosan polysulfate); phenylbutazone (e.g., phenylbutazone sodium glycerate); pirfenidone; piroxicam (e.g., piroxicam cinnamate, piroxicam ethanolamine); pirprofen; penazate; prifelone; (ii) propandolic acid; proquinolizone; propiconazole (e.g., propiconazole citrate); rimexolone; 2, chloromazali; (ii) willowherb leshi; salicylic acids (e.g., salicylic acid); a salt of a compound of Sanaretidine; salsalate; sanguinarine (e.g., sanguinarine chloride); -clazonone; silketeine; sudoxicam; sulindac; suprofen; tamicin; talniflumate; talosulfate; terbufuron; tenidapa (e.g., tenidapa sodium); tenoxicam; tixicam; a teximide; tetrahydroindamide; sulfuric acid; ticortisone (e.g., ticortisone pivalate); tolmetin (e.g., tolmetin sodium); (ii) triclosan; a trifluorourethane; zidometacin; and zomepirac (e.g., sodium zomepirac).

Examples of suitable antiproliferative agents include, but are not limited to, acivicin; aclarubicin; aristodazole (e.g., aristodazole hydrochloride); (ii) abelmoscine; doxorubicin; (ii) Alexanox; aldesleukin; altretamine; an apramycin; amenthraquinone (e.g., amenthraquinone acetate); aminoglutethimide; amsacrine; anastrozole; an atramycin; an asparaginase enzyme; a triptyline; azacitidine; azatepa; (ii) azomycin; batimastat; benztepa; bicalutamide; bisantrene (e.g., bisantrene hydrochloride); bis-naphthalene methods (e.g., bis-naphthalene method of disulfonic acid); bizelesin; bleomycin (e.g., bleomycin sulfate); brequinar (e.g., brequinar sodium); briprimine; busulfan; actinomycin C; (ii) carpoterone; a carbimide; a carbapenem; carboplatin; carmustine; carrubicin (e.g., carrubicin hydrochloride); folding to get new; cediogo; chlorambucil; a sirolimus; cisplatin; cladribine; carbetocin phosphate A-4; clinatol (e.g., clinatol mesylate); cyclophosphamide; cytarabine; dacarbazine; actinomycin D; daunorubicin (e.g., daunorubicin hydrochloride); decitabine; (ii) dexomaplatin; dizyguanine (e.g., dizyguanine mesylate); diazaquinone; docetaxel; doxorubicin (e.g., doxorubicin hydrochloride); droloxifene (e.g., droloxifene citrate); drotaandrosterone (e.g., drotaandrosterone propionate); daptomycin; edatrexae; eflornithine (e.g., eflornithine hydrochloride); elsamitrucin; enloplatin; an enpu urethane; epinastine; epirubicin (e.g., epirubicin hydrochloride); (ii) ebuzole; isosbacin (e.g., isosbacin hydrochloride); estramustine (e.g., estramustine sodium phosphate); etanidazole; etoposide (e.g., etoposide phosphate); etophenine; fadrozole (e.g., fadrozole hydrochloride); fazarabine; fenretinide; floxuridine; fludarabine (e.g., fludarabine phosphate); fluorouracil; (iii) flucitabine; a phosphorus quinolone; fostrexasin (e.g., fostrexasin sodium); gemcitabine (e.g., gemcitabine hydrochloride); a hydroxyurea; idarubicin (e.g., idarubicin hydrochloride); ifosfamide; ilofovir dipivoxil; interferon alpha-2 a; interferon alpha-2 b; interferon alpha-n 1; interferon alpha-n 3; interferon beta-Ia; interferon gamma-Ib; iproplatin; irinotecan (e.g., irinotecan hydrochloride); lanreotide (e.g., lanreotide acetate); letrozole; leuprolide (e.g., leuprolide acetate); liazole (e.g., liazole hydrochloride); lometrexol (e.g., sodium lometrexol); lomustine; losoxanthraquinone (e.g., losoxanthraquinone hydrochloride); (ii) maxolone; maytansine; nitrogen mustards (e.g., mechlorethamine hydrochloride); megestrol (e.g., megestrol acetate); melengestrol (e.g., melengestrol acetate); melphalan; (ii) a melanoril; mercaptopurine; methotrexate (e.g., sodium methotrexate); chlorpheniramine; meltupipide; mitodomide; mitokacin; mitorubin; mitoxantrone; mitosin; mitomycin; mitospirane culturing; mitotane; mitoxantrone (e.g., mitoxantrone hydrochloride); mycophenolic acid; nocodazole; a noggin; (ii) orelbulin; ormaplatin; oshuzuren; paclitaxel; a pemetrexed; a calicheamicin; nemadectin; pellomycin (e.g., pellomycin sulfate); cultivating phosphoramide; pipobroman; piposulfan; piroanthraquinone (e.g., piroanthraquinone hydrochloride); (ii) a plicamycin; pramipexole; porfimer (e.g., porfimer sodium); porphyrins; deltemustine; procarbazine (e.g., procarbazine hydrochloride); puromycin (e.g., puromycin hydrochloride); pyrazole furan rhzomorph; (ii) lybodenosine; pirimid; safrog (e.g., safrog hydrochloride); semustine; octreozine; sparfosate (e.g., sparfosate sodium); sparsomycin; germanospiramine (e.g., germanospiramine hydrochloride); spiromustine; spiroplatinum; streptonigrin; streptozotocin; a sulfochlorophenylurea; a talithromycin; tekrainian (e.g., tekrainian sodium); tegafur; tiloxanthraquinone (e.g., tiloxanthraquinone hydrochloride); temoporfin; (ii) teniposide; a tiroxiron; a testosterone ester; (ii) a thiopurine; sulfur guanine; thiotepa; (ii) a thiazole carboxamide nucleoside; tirapazamine; topotecan (e.g., topotecan hydrochloride); toremifene (e.g., toremifene citrate); a tropilone (e.g., tropilone acetate); triciribine (e.g., triciribine phosphate); trimetrexate (e.g., glucuronic acid trimetrexate); triptorelin; tobramzole (e.g., tobramzole hydrochloride); uracil mustard; uretipi; vapreotide; verteporfin; vinblastine; vincristine (e.g., vincristine sulfate); vindesine (e.g., vindesine sulfate); vincepidine; vinblastin ester; vinblastine; vinorelbine (e.g., vinorelbine tartrate); vinclodine; vinzolidine; (ii) vorozole; zeniplatin; 1, neat setastine; and zorubicin (e.g., zorubicin hydrochloride). Additional anti-cancer Agents include those disclosed in "The pharmaceutical Basis of Therapeutics", eighth edition, 1990, McGraw-Hill, inc. (Health services Division), chapter 52 anti-neoplastic Agents (Paul cabarestri and Bruce a. chamner), and The introduction thereof, page 1202 and 1263, by Goodman and Gilman, The contents of which are incorporated herein by reference.

Examples of therapeutically active agents suitable for inclusion in liposomes according to some embodiments described herein (e.g., as molecules or moieties of pharmaceutical agents) include, but are not limited to, amphotericin B, cisplatin, cytarabine, daunorubicin, doxorubicin, influenza hemagglutinin and/or neuraminidase, morphine, surfactant protein B, surfactant protein C, verteporfin, and vincristine.

Examples of medical conditions that can be treated by amphotericin B (e.g., by injection and/or infusion of amphotericin B-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, protozoal infections (e.g., leishmaniasis) and fungal infections, such as aspergillosis, blastomycosis, candidiasis, coccidioidomycosis, and cryptococcosis.

Inclusion of amphotericin B in liposomes according to any of the respective embodiments described herein can optionally result in reduced toxicity (e.g., reduced renal toxicity) and/or improved pharmacokinetics (e.g., in the treatment of central nervous system infections).

Examples of medical conditions that can be treated by cytarabine (e.g., by injection and/or infusion of cytarabine-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, cancers, such as leukemias (e.g., acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia) and lymphomas (e.g., non-hodgkin's lymphoma, lymphomatous meningitis).

Inclusion of cytarabine in liposomes according to any of the respective embodiments described herein may optionally result in enhanced efficacy (e.g., in the treatment of lymphoma meningitis).

Examples of medical conditions that can be treated by cisplatin (e.g., by injection and/or infusion of cisplatin-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, cancers, such as bladder cancer, brain tumors, breast cancer, cervical cancer, esophageal cancer, germ cell tumors, head and neck cancer, lung cancer (e.g., small cell lung cancer), mesothelioma, neuroblastoma, ovarian cancer, pancreatic cancer, testicular cancer, and sarcoma.

Examples of medical conditions that can be treated by daunorubicin (e.g., by injection and/or infusion of daunorubicin-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, cancers, such as leukemias (e.g., acute myelogenous leukemia, acute lymphocytic leukemia, chronic myelogenous leukemia), lymphomas (non-hodgkin's lymphoma), neuroblastomas, and kaposi's sarcoma (e.g., AIDS-related kaposi's sarcoma).

Inclusion of daunorubicin in liposomes according to any of the respective embodiments described herein can optionally result in improved pharmacokinetic profiles (e.g., increased concentrations) at the site of the injury (e.g., kaposi's sarcoma injury).

Examples of medical conditions that can be treated by doxorubicin (e.g., by injection and/or infusion of liposomes containing doxorubicin according to any of the respective embodiments described herein) include, but are not limited to, cancers, such as breast cancer (e.g., in combination with cyclophosphamide), bladder cancer, kaposi's sarcoma (such as AIDS-related kaposi's sarcoma), leukemia (such as acute lymphocytic leukemia), lymphoma (hodgkin's lymphoma, multiple myeloma), lung cancer, ovarian cancer, soft tissue sarcoma, gastric cancer, and thyroid cancer.

Inclusion of doxorubicin in liposomes according to any of the respective embodiments described herein can optionally result in reduced toxicity (e.g., cardiotoxicity) and/or improved pharmacokinetic profiles (e.g., increased concentration) at the site of the injury (e.g., kaposi's sarcoma injury).

Examples of medical conditions that can be treated by estradiol (e.g., by topical administration of estradiol-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, menopause, hypogonadism, sex anxiety, infertility, lactation, hyperphagia (e.g., in adolescent girls), and hormone sensitive cancers (e.g., breast cancer, prostate cancer).

Influenza hemagglutinin and/or neuraminidase can optionally be used to treat (e.g., prevent) influenza, for example, as an influenza vaccine. The vaccine may optionally be administered by injection and/or infusion of a liposome comprising hemagglutinin and/or neuraminidase according to any of the respective embodiments described herein (optionally in the form of viral particles).

Examples of medical conditions that can be treated by morphine (e.g., by injection and/or infusion of morphine-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, acute and chronic pain (e.g., pain associated with surgery, myocardial infarction, and/or labor) and shortness of breath.

Inclusion of morphine in a liposome according to any of the respective embodiments described herein can optionally result in an extension (e.g., at least 48 hours) of the time period for which therapeutically effective levels of morphine are present in the blood of the subject.

Respiratory distress syndrome (e.g., in a preterm infant) is a non-limiting example of a medical condition that can be treated by surfactant protein B and/or surfactant protein C (e.g., by pulmonary (e.g., intratracheal) administration of surfactant protein B and/or C-containing liposomes according to any of the respective embodiments described herein).

Examples of medical conditions that can be treated by verteporfin (e.g., by injection and/or infusion of verteporfin-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, ocular diseases, such as abnormal blood vessels in the eye (e.g., associated with wet and/or age-related macular degeneration), pathological myopia, ocular histoplasmosis, and central serous retinopathy, such as associated with photodynamic therapy (e.g., where verteporfin is administered prior to laser treatment).

Examples of medical conditions that can be treated by vincristine (e.g., by injection and/or infusion of vincristine-containing liposomes according to any of the respective embodiments described herein) include, but are not limited to, thrombocytopenic purpura (e.g., thrombotic thrombocytopenic purpura or idiopathic thrombocytopenic purpura) and cancer, such as neuroblastoma, wilms ' tumor, melanoma, lung cancer (e.g., small cell lung cancer), leukemia (e.g., acute myeloid leukemia, acute lymphocytic leukemia), and lymphoma (e.g., hodgkin's lymphoma or non-hodgkin's lymphoma).

Inclusion of vincristine in the liposomes according to any of the respective embodiments described herein may optionally result in enhanced efficacy, enhanced entry of vincristine into cells, increased plasma concentration and/or circulating life of vincristine, and/or reduced toxicity (e.g., reduced neurotoxicity).

The skilled person will be able to readily determine which medical condition may be treated by a given therapeutically active agent, and which therapeutically active agent may be suitable for treating a given medical condition.

In some embodiments of any of the embodiments described herein, the liposome is for use in treating a proliferative disease or disorder (e.g., cancer), and the therapeutically active agent is an antiproliferative agent according to any of the respective embodiments described herein.

In some embodiments of any of the embodiments described herein, the liposome is for use in treating an inflammatory disease or disorder (e.g., cancer), and the therapeutically active agent is an analgesic and/or an anti-inflammatory agent according to any of the respective embodiments described herein.

In some embodiments of any of the embodiments described herein, the liposome is used to treat a synovial joint disorder (e.g., by systemic and/or intra-articular administration), optionally an inflammatory synovial joint disorder. Examples of synovial joint conditions that can be treated according to embodiments of the invention include, but are not limited to, arthritis (e.g., osteoarthritis, rheumatoid arthritis and/or psoriatic arthritis), bursitis, carpal tunnel syndrome, fibromyositis, gout, locked joints (e.g., locked joints associated with osteochondritis dissecans and/or synovial chondromatosis), tendonitis, traumatic joint injury and joint damage associated with surgery.

The joint damage associated with surgery may optionally be associated with surgery that causes damage directly to the joint surface (e.g., through an incision) and/or surgery that only indirectly damages the joint surface. For example, surgery to repair or otherwise affect tissue (e.g., ligaments and/or menisci) near a joint may be associated with joint damage due to mechanical changes in the joint.

Traumatic joint injury can optionally be injury caused directly by trauma (e.g., caused at the time of trauma) and/or injury caused by a previous trauma (e.g., post-traumatic injury occurring at some time after trauma).

Formulation and administration:

the liposomes according to any of the embodiments described herein can be administered to a subject by themselves or as part of a pharmaceutical composition.

As used herein, "pharmaceutical composition" refers to a formulation of one or more active ingredients described herein with other chemical components such as pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration of the active ingredient (e.g., a therapeutically active agent with or without liposomes according to any of the respective embodiments described herein) to a subject.

Throughout this document, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to a subject when administered in the intended manner, and does not abrogate the activity and properties of the liposomes in the composition (e.g., their ability to reduce the coefficient of friction of a surface, as described herein in any of the respective embodiments). Non-limiting examples of carriers are: propylene glycol, saline, emulsions and mixtures of organic solvents with water, as well as solids (e.g., powders) and gaseous carriers.

In some embodiments of any of the embodiments described herein, the composition comprises an aqueous carrier, which is a pharmaceutically acceptable carrier, e.g., wherein the composition is a solution.

Herein, the term "excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of the active ingredient of the present invention or to increase shelf-life stability. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and salts and various types of starches, cellulose derivatives, gelatin, vegetable oils, EDTA, EGTA, poly-L-lysine, polyethyleneimine, pimecronium bromide, polyethylene glycol, and other polyanions or monoanions. The pharmaceutical composition may advantageously take the form of a foam, aerosol or gel.

In some embodiments of any of the respective embodiments described herein, the pharmaceutical composition further comprises a water-soluble biopolymer, such as a polypeptide and/or a polysaccharide. The polymer may be ionic or non-ionic.

As used herein, the phrase "water-soluble biopolymer" includes biopolymers having a solubility of at least 1 g/l in an aqueous (e.g., water) environment at pH7 (at 25 ℃).

In some embodiments of any of the embodiments described herein, the water-soluble polymer has a solubility (under the conditions described above) of at least 2 grams per liter. In some embodiments, the solubility is at least 5 grams per liter. In some embodiments, the solubility is at least 10 grams per liter. In some embodiments, the solubility is at least 20 grams per liter. In some embodiments, the solubility is at least 50 grams per liter. In some embodiments, the solubility is at least 100 grams per liter.

Examples of suitable nonionic water-soluble polymers include, but are not limited to, polyvinylpyrrolidone (also interchangeably referred to herein as povidone and/or PVP) and polyethylene oxide (also interchangeably referred to herein as PEO, PEG, and/or polyethylene glycol).

Hyaluronic acid (ionic polysaccharide) is an exemplary biopolymer.

As exemplified herein, water-soluble polymers can enhance the in vivo retention of liposomes according to some embodiments described herein.

Pharmaceutical compositions (e.g., liposome solutions) for use in accordance with the present invention may be formulated in a conventional manner (e.g., by conventional mixing or dissolution procedures) using one or more pharmaceutically acceptable carriers (optionally including excipients and adjuvants) that facilitate processing of the liposomes (and optionally also the water-soluble polymers described herein) into a formulation that may be used pharmaceutically. The appropriate formulation depends on the route of administration chosen.

Techniques for formulating and administering compounds can be found in "Remington's Pharmaceutical Sciences" Mack Publishing co., Easton, PA, latest edition, which is incorporated herein by reference.

The liposomes described herein (optionally together with the water-soluble polymers described herein) can be formulated as aqueous solutions themselves. Additionally, the solution may be in the form of a suspension and/or emulsion (e.g., an aqueous phase of a suspension or a water-in-oil, oil-in-water, or water-in-oil emulsion), for example, to increase the viscosity of the formulation. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, for example sodium carboxymethyl cellulose, sorbitol or dextran. Optionally, the suspension may also contain suitable stabilizers or agents that increase the solubility of the liposomes described herein (and/or the optional water-soluble polymers described herein), e.g., to allow for the preparation of highly concentrated solutions.

In some embodiments, the liposomes described herein (optionally together with the water-soluble polymers described herein) can be in powder form for constitution with a suitable vehicle (e.g., sterile, pyrogen-free water) before use.

Suitable routes of administration include any of a variety of suitable systemic and/or local routes of administration.

Suitable routes of administration may include, for example, inhalation, oral, buccal, rectal, transmucosal, topical, transdermal, intradermal, nasal, intestinal and/or parenteral routes; intramuscular, subcutaneous and/or intramedullary routes of injection; intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal and/or intraocular injection routes, catheterization with or without vascular balloons; and/or direct injection into a tissue region of a subject.

Parenteral administration according to any of the embodiments described herein can optionally be systemic (e.g., intravenous) and/or local (e.g., intra-articular). The liposomes described herein (optionally with a water-soluble polymer as described herein) can be formulated for parenteral administration, for example by bolus injection or continuous infusion. The compositions may be in the form of suspensions, solutions (e.g., aqueous solutions of the active ingredient in water-soluble form), or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate, triglycerides or liposomes.

For injection, the liposomes described herein (optionally with a water-soluble polymer as described herein) can be formulated in aqueous solution, preferably in a physiologically compatible buffer, such as Hank's solution, Ringer's solution, histidine buffer, or physiological saline buffer, with or without an organic solvent (e.g., propylene glycol, polyethylene glycol).

Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, optionally with an added preservative.

For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

For oral administration, pharmaceutical compositions can be readily formulated by combining the active ingredient with pharmaceutically acceptable carriers well known in the art. These carriers enable the pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient. Pharmaceutical preparations for oral use can be prepared using solid excipients, optionally grinding the resulting mixture, and processing the mixture of granules, if desired after adding suitable auxiliaries, to obtain tablets or dragee cores. Suitable excipients are in particular fillers, for example sugars, including lactose, sucrose, mannitol or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose; and/or physiologically acceptable polymers, such as polyvinylpyrrolidone (PVP). If desired, disintegrating agents can be added, such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof, such as sodium alginate.

Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbomer gel, polyethylene glycol, titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyes or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active ingredient doses.

Pharmaceutical compositions for oral use include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Push-fit capsules can contain the active ingredient in admixture with filler (e.g., lactose), binder (e.g., starch), lubricant (e.g., talc or magnesium stearate), and optional stabilizers. In soft capsules, the active ingredient may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added.

For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

For administration by the inhalation route, the active ingredients used in accordance with the present invention may be delivered in aerosol/spray presentation form from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g., a chlorofluorocarbon such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane; carbon dioxide; or a volatile hydrocarbon such as butane, propane, isobutane or mixtures thereof. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin (cartridges) containing a powder mix of the active ingredient and a suitable powder base such as lactose or starch may be formulated for use in the dispenser.

The pharmaceutical compositions may also be formulated in rectal compositions such as suppositories or retention enemas, using, for example, conventional suppository bases such as cocoa butter or other glycerides.

The composition can be formulated wherein the liposomes are included in an amount effective to achieve the intended purpose, e.g., an amount effective to prevent, alleviate, or ameliorate symptoms of the disorder in the subject being treated.

The dosage may vary depending on the dosage form employed, the route of administration utilized, and the location of administration (e.g., the volume and/or surface of the area in contact with the liposomes).

Determination of a therapeutically effective amount is well within the capability of those skilled in the art (e.g., based on what is known in the art for any given therapeutically active agent), particularly in light of the detailed disclosure provided herein.

For any formulation used in the methods of the invention, a therapeutically effective amount or dose can be estimated initially from in vitro and cell culture and in vivo assays. For example, a dose can be formulated in animal models to achieve a desired concentration or titer. This information can be used to more accurately determine useful doses in humans.

Toxicity and/or therapeutic efficacy of the active ingredients described herein can be determined by standard pharmaceutical procedures in vitro, in cell cultures or experimental animals. The data obtained from these in vitro and cell culture assays and animal studies can be used to formulate a range of dosages for use in humans. The dosage may vary depending on the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage may be selected by the individual physician in accordance with the circumstances of the patient. (see for example the Fingl @ system,et al1975, see "The Pharmacological Basis of Therapeutics", Ch. 1 p.1).

The dosage amounts and intervals may be adjusted individually to provide plasma or brain levels (minimum effective concentration, MEC) of the active ingredient(s) sufficient to achieve the desired therapeutic effect. The MEC for each formulation will vary, but can be estimated from in vitro data. The dose required to achieve MEC depends on the individual characteristics and the route of administration. Detection assays can be used to determine plasma concentrations.

The amount of composition administered will, of course, depend on the subject being treated, the severity of the affliction, the mode of administration, the judgment of the prescribing physician, and the like.

Depending on the severity and responsiveness of the condition to be treated, administration may be a single or multiple administrations, with the course of treatment lasting from days to weeks or until cure is effected or remission of the disease state is achieved.

If desired, compositions according to embodiments of the invention (e.g., liposome solutions) can be presented in a packaging or dispenser device, such as an FDA approved kit, which can contain one or more unit dosage forms containing one or more active ingredients (e.g., liposomes as described herein). The package may for example comprise a metal or plastic foil, such as but not limited to a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The package or dispenser may also be accompanied by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice reflects approval by the agency of the form of the composition for human or veterinary administration. For example, such a notification may be a label approved by the U.S. food and drug administration for a prescription drug or an approved product insert. Compositions comprising liposomes (optionally in conjunction with a water-soluble polymer as described herein) as described herein in any of the respective embodiments, formulated in a pharmaceutically acceptable carrier can also be prepared, placed in a suitable container, and labeled for treatment of an indicated condition or diagnosis, as detailed herein.

Other definitions:

herein, the term "hydrocarbon" describes an organic moiety comprising as its basic skeleton a chain of carbon atoms predominantly substituted with hydrogen atoms. The hydrocarbons may be saturated or unsaturated, contain aliphatic, alicyclic, or aromatic moieties, and may be optionally substituted with one or more substituents (other than hydrogen). The substituted hydrocarbon may have one or more substituents, wherein each substituent may independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, oxo, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine. The hydrocarbon may be a terminal group or a linking group, as these terms are defined herein. The hydrocarbon moiety is optionally interrupted by one or more heteroatoms including, but not limited to, one or more oxygen, nitrogen and/or sulfur atoms. In some embodiments of any of the embodiments described herein involving a hydrocarbon, the hydrocarbon is not interrupted by any heteroatom.

Preferably, the hydrocarbon moiety has from 1 to 20 carbon atoms. Whenever a numerical range such as "1 to 20" is stated herein, it is implied that the group, in this case alkyl, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, and the like, up to and including 20 carbon atoms.

Herein, the term "alkyl" describes a saturated aliphatic hydrocarbon end group as defined herein, including straight and branched chain groups. Preferably, the alkyl group has 1 to 20 carbon atoms. More preferably, the alkyl group is a medium size alkyl group having 1 to 10 carbon atoms. Most preferably, unless otherwise specified, alkyl is lower alkyl having 1 to 4 carbon atoms. The alkyl group may be substituted or unsubstituted. The substituted alkyl group may have one or more substituents, wherein each substituent may independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.

The term "alkylene" describes a saturated aliphatic hydrocarbon linking group, as that term is defined herein, which differs from alkyl as defined herein only in that alkylene is a linking group and not a terminal group.

Herein, the term "alkenyl" describes an unsaturated aliphatic hydrocarbon end group comprising at least one carbon-carbon double bond, including straight and branched chain groups. Preferably, the alkenyl group has 2 to 20 carbon atoms. More preferably, the alkenyl group is a medium size alkenyl group having 2 to 10 carbon atoms. Most preferably, unless otherwise specified, alkenyl is lower alkenyl having 2 to 4 carbon atoms. The alkenyl group may be substituted or unsubstituted. The substituted alkenyl group may have one or more substituents, wherein each substituent may independently be, for example, cycloalkyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.

Herein, the term "alkynyl" describes an unsaturated aliphatic hydrocarbon end group comprising at least one carbon-carbon triple bond, including straight and branched chain groups. Preferably, the alkynyl group has 2 to 20 carbon atoms. More preferably, the alkynyl group is a medium size alkynyl group having 2 to 10 carbon atoms. Most preferably, unless otherwise specified, alkynyl is lower alkynyl having 2 to 4 carbon atoms. The alkynyl group may be substituted or unsubstituted. The substituted alkynyl group may have one or more substituents, wherein each substituent may independently be, for example, cycloalkyl, alkenyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine.

The term "cycloalkyl" describes an all-carbon monocyclic or fused-ring (i.e., rings that share adjacent pairs of carbon atoms) group in which one or more of the rings does not have a completely conjugated pi-electron system. Cycloalkyl groups may be substituted or unsubstituted. The substituted cycloalkyl group may have one or more substituents, wherein each substituent may independently be, for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine. Cycloalkyl groups may be an end group, as that phrase is defined herein, wherein the end group is attached to a single adjacent atom or is a linking group that connects two or more moieties, as that phrase is defined herein.

The term "aryl" describes an all-carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) end group having a completely conjugated pi-electron system (as that term is defined herein). The aryl group may be substituted or unsubstituted. The substituted aryl group may have one or more substituents, wherein each substituent may independently be, for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine. Phenyl and naphthyl are representative aryl end groups.

The term "heteroaryl" describes a monocyclic or fused ring (i.e., rings that share adjacent pairs of atoms) group having one or more atoms, such as nitrogen, oxygen, and sulfur, in one or more rings, and further having a fully conjugated pi-electron system. Examples of heteroaryl groups include, but are not limited to, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, and purine. Heteroaryl groups may be substituted or unsubstituted. The substituted heteroaryl group may have one or more substituents, wherein each substituent may independently be, for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine. Heteroaryl groups can be an end group, as that phrase is defined herein, wherein the end group is attached to a single adjacent atom, or a linking group that connects two or more moieties, as that phrase is defined herein. Representative examples are pyridine, pyrrole, oxazole, indole, purine and the like.

The term "arylene" describes a single or fused ring polycyclic linking group as that term is defined herein and includes linking groups other than aryl or heteroaryl as those groups are defined herein except that arylene is a linking group and not an end group.

The term "heteroalicyclic" describes a monocyclic or fused ring group having one or more atoms in one or more rings, such as nitrogen, oxygen, and sulfur. The rings may also have one or more double bonds. However, rings do not have a completely conjugated pi-electron system. The heteroalicyclic group may be substituted or unsubstituted. The substituted heteroalicyclic group can have one or more substituents, wherein each substituent can independently be, for example, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, azide, sulfonamide, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, and hydrazine. A heteroalicyclic group can be an end group, as that phrase is defined herein, wherein it is attached to a single adjacent atom, or a linking group that connects two or more moieties, as that phrase is defined herein. Representative examples are piperidine, piperazine, tetrahydrofuran, tetrahydropyran, morpholine and the like.

As used herein, the terms "amine" and "amino" describe both the-NRxRy end group and the-NRx-linking group, wherein Rx and Ry are each independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or heteroalicyclic, as these terms are defined herein. When Rx or Ry is heteroaryl or heteroalicyclic, the amine nitrogen atom is bonded to a carbon atom of the heteroaryl or heteroalicyclic ring. The carbon atom attached to the nitrogen atom of the amine is not substituted with = O or = S, and in some embodiments, is not substituted with any heteroatom.

Thus, the amine group may be a primary amine, wherein Rx and Ry are both hydrogen; a secondary amine, wherein Rx is hydrogen and Ry is alkyl, cycloalkyl, aryl, heteroaryl, or heteroalicyclic; or a tertiary amine, wherein Rx and Ry are each independently alkyl, cycloalkyl, aryl, heteroaryl, or heteroalicyclic.

The terms "hydroxy" and "hydroxyl" describe the-OH group.

The term "alkoxy" describes both-O-alkyl and-O-cycloalkyl end groups, or-O-alkylene or-O-cycloalkyl linking groups, as defined herein.

The term "aryloxy" describes both the-O-aryl and-O-heteroaryl end groups, or the-O-arylene-linking group, as defined herein.

The term "thiol" describes an-SH group.

The term "thioalkoxy" describes both an-S-alkyl and an-S-cycloalkyl end group, or an-S-alkylene or-S-cycloalkyl linking group, as defined herein.

The term "thioaryloxy" describes both-S-aryl and-S-heteroaryl end groups, or-S-arylene-linking groups, as defined herein.

The terms "cyano" and "nitrile" describe a-C ≡ N group.

The term "nitro" describes-NO₂A group.

The term "oxo" describes an = O group.

The term "azide" describes-N = N⁺=N^-A group.

The term "azo" describes an-N = N-Rx end group or-N = a linking group, wherein Rx is as defined herein.

The terms "halide" and "halo" refer to fluorine, chlorine, bromine or iodine.

The term "phosphate ester" means-O-P (= O) (ORx) -OR_YTerminal group, or means an-O-P (= O) (ORx) -O-linking group, wherein Rx and R_YAs defined herein.

The terms "phosphono" and "phosphonate" refer to-P (= O) (ORx) -OR_YTerminal group, or means a-P (= O) (ORx) -O-linking group, wherein Rx and R_YAs defined herein.

The term "phosphinyl" refers to-PRxR_YGroup, wherein Rx and R_YAs defined above.

The term "sulfoxide" or "sulfinyl" describes a-S (= O) -Rx end group or-S (= O) -linker group, wherein Rx is as defined herein.

The terms "sulfonate" and "sulfonyl" describe-S (= O)₂-Rx terminal group or-S (= O)₂-a linking group, wherein Rx is as defined herein.

The terms "sulfonamide" and "sulfonamido" as used herein include both S-sulfonamide and N-sulfonamide terminal groups, and-S (= O)₂-NRx-linking group.

The term "S-sulfonamide" describes-S (= O)₂-NRxR_YEnd groups, wherein Rx and R_YAs defined herein.

The term "N-sulfonamide" describes RxS (= O)₂–NR_Y-end groups, wherein Rx and R_YAs defined herein.

The term "carbonyl" as used herein describes a-C (= O) -Rx end group or a-C (= O) -linking group, wherein Rx is as defined herein.

The term "acyl" as used herein describes a-C (= O) -Rx end group, wherein Rx is as defined herein.

The term "thiocarbonyl" as used herein describes a-C (= S) -Rx end group or a-C (= S) -linking group, wherein Rx is as defined herein.

The terms "carboxy" and "carboxyl" as used herein include both C-and O-carboxy end groups, and a-C (= O) -O-linking group.

The term "C-carboxy" describes a-C (= O) -ORx end group, wherein Rx is as defined herein.

The term "O-carboxy" describes the-OC (= O) -Rx end groups, wherein Rx is as defined herein.

The term "urea" describes a-NRxC (= O) -NRyRw end group or-NRxC (= O) -NRy-linking group, where Rx and Ry are as defined herein and Rw is as defined herein for Rx and Ry.

The term "thiourea" describes the-NRx-C (= S) -NRyRw end group or the-NRx-C (= S) -NRy-linking group, where Rx, Ry and Ry are as defined herein.

The terms "amide" and "amido" as used herein include both C-amide and N-amide end groups, and a-C (= O) -NRx-linking group.

The term "C-amide" describes a-C (= O) -NRxRy end group, wherein Rx and Ry are as defined herein.

The term "N-amide" describes RxC (= O) -NRy-end groups, where Rx and Ry are as defined herein.

The term "carbamoyl" or "carbamate" as used herein includes N-carbamate and O-carbamate end groups, and an-OC (= O) -NRx-linking group.

The term "N-carbamate" describes the RyOC (= O) -NRx-end group, where Rx and Ry are as defined herein.

The term "O-carbamate" describes the-OC (= O) -NRxRy end group, where Rx and Ry are as defined herein.

The term "thiocarbamoyl" or "thiocarbamate" as used herein includes O-thiocarbamate, S-thiocarbamate, and N-thiocarbamate terminal groups, and-OC (= S) -NRx-or-SC (= O) -NRx-linking groups.

The terms "O-thiocarbamate" and "O-thiocarbamoyl" describe an-OC (= S) -NRxRy end group, where Rx and Ry are as defined herein.

The terms "S-thiocarbamate" and "S-thiocarbamoyl" describe the-SC (= O) -NRxRy end group, where Rx and Ry are as defined herein.

The terms "N-thiocarbamate" and "N-thiocarbamoyl" describe either the RyOC (= S) NRx-or RySC (= O) NRx-end group, where Rx and Ry are as defined herein.

The term "guanidine" describes a-RxNC (= N) -NRyRw terminal group or a-RxNC (= N) -NRy-linking group, where Rx, Ry, and Rw are as defined herein.

As used herein, the term "hydrazine" describes a-NRx-NRyRw terminal group or-NRx-NRy-linking group, wherein Rx, Ry and Rw are as defined herein.

As used herein, the term "about" means ± 10%, and optionally ± 5%.

The terms "comprising," including, "" having, "and their equivalents mean" including but not limited to.

The term "consisting of means" including and limited to.

The term "consisting essentially of means that the composition, method, or structure may include other ingredients, steps, and/or portions, provided that the other ingredients, steps, and/or portions do not materially alter the basic and novel characteristics of the claimed composition, method, or structure.

As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

Throughout this application, various embodiments of the present invention may be presented in a range format. It is to be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, description of a range such as 1 to 6 should be considered to specifically disclose sub-ranges, e.g., 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as individual numbers within that range, e.g., 1,2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is intended to include any number of the referenced number (fractional or integer) within the indicated range. The phrases "range between a first indicated number and a second indicated number" and "range/range" of the first indicated number "to the second indicated number are used interchangeably herein and are meant to include the first and second indicated numbers and all fractional and integer numbers therebetween.

As used herein, the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by, practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

As used herein, the term "treating" includes eliminating, substantially inhibiting, slowing or reversing the progression of the disorder, substantially ameliorating clinical or aesthetic symptoms of the disorder or substantially preventing the appearance of clinical or aesthetic symptoms of the disorder.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments should not be considered essential features of those embodiments, unless the embodiment does not function without those elements.

Various embodiments and aspects of the present invention as described above and as claimed in the appended claims section find experimental support in the following examples.

Examples

Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non-limiting manner.

Materials and methods

Materials:

2-Bromoisobutyryl bromide was obtained from Sigma-Aldrich.

Chloroform was obtained from Sigma-Aldrich.

CuBr was obtained from Sigma-Aldrich.

DiR (1,1' -dioctadecyl-3, 3,3',3' -tetramethylindotricarbocyanine iodide) is obtained from Molecular Probes.

Distearoyl phosphatidylethanolamine (DSPE) is obtained from Avanti Polar Lipids.

Distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG, PEG Mw of 2000Da (DSPE-PEG 2000) or 550Da (DSPE-PEG 550)) was obtained from Avanti.

Hyaluronic acid (2500 had) was obtained from Creative pegweights.

Hydrogenated Soybean Phosphatidylcholine (HSPC) is obtained from Lipoid GmbH.

Methanol was obtained from Bio-Lab.

N,N,N',N",N"Pentamethyldiethylenetriamine (PMDETA) was obtained from Sigma-Aldrich.

O- (2-Methacryloyloxyethyl) Phosphorylcholine (MPC) was obtained from biocompatable Corporation (UK).

Phosphate Buffered Saline (PBS) was obtained from Sigma-Aldrich.

The water was purified using a Barnstead NanoPure system to a resistance of 18.2M Ω cm with a total organic content level < about l ppb.

Synthesis of DSPE-PMPC:

as illustrated in scheme 1Described, using the procedure described in international patent application IL2016/051372, phospholipids with polymeric phosphorylcholine derivatives were prepared from the phospholipids DSPE (distearoylphosphatidylethanolamine) and the phosphorylcholine derivative MPC (O- (2-methacryloyloxyethyl) phosphorylcholine). Briefly, DSPE was reacted with 2-bromoisobutyryl bromide to obtain a free radical initiator (DSPE-Br) which was then used in atom transfer radical polymerization with MPC (O- (2-methacryloyloxyethyl) phosphorylcholine) at 60 ℃ in the presence of CuBr and PMDETA (N, N', N "-pentamethyldiethylenetriamine). The DSPE-PMPC obtained is purified by dialysis and passed¹The H-NMR spectrum determined a pMPC fraction with a Mw of about 2 kDa.

Scheme 1

Preparing liposome:

to prepare liposomes, DSPE-PMPC (prepared as described above) and HSPC were dissolved in methanol and chloroform (2ml, 1:1 v/v) at a molar ratio of 2:98(DSPE-PMPC: HSPC). The organic solvent was then dried overnight with nitrogen to form a dry film. Multilamellar vesicles (MLVs) are then prepared by hydrating lipids in PBS (phosphate buffered saline) at least 5 ℃ above the melting point of the lipids, followed by sonication. To prepare pegylated liposomes, the above procedure was applied to the following mixture: DSPE-PEG2000 and HSPC and DSPE-PEG550 and HSPC, both at a molar ratio of 2:98, to form MLV. The organic solvent used was chloroform for HSPC and methanol for DSPE-PMPC and DSPE-PEG.

MLV was scaled down to form Large Unilamellar Vesicles (LUV) by stepwise extrusion through polycarbonate membranes using a Lipex 10ml extruder system (Northern Lipids, Canada). In some cases, MLVs were scaled down to form Small Unilamellar Vesicles (SUVs) by stepwise extrusion through polycarbonate membranes using a Lipex 10ml extruder system (Northern Lipids, Canada), starting with 400nm pore size membranes and ending with 50nm pore size membranes.

Dynamic light scattering measurements (DLS) showed that the mean diameter of the LUV was about 170nm and the mean diameter of the SUV was about 80 nm.

Animal experiments:

all Animal experiments were performed under the protocol approved by the Institutional Animal Care and Use Committee (IACUC) of Weizmann Institute of Science, Israel, according to the National Institute of Health (NIH) Animal research guidelines.