DRUG DELIVERY POLYMERS AND USES THEREOF
20170348431 · 2017-12-07
Inventors
- Jeremiah A. Johnson (Boston, MA)
- Longyan Liao (Cambridge, MA)
- Jonathan Christopher Barnes (Waltham, MA)
Cpc classification
A61K31/704
HUMAN NECESSITIES
A61K33/00
HUMAN NECESSITIES
C08G69/48
CHEMISTRY; METALLURGY
A61K31/4745
HUMAN NECESSITIES
A61K47/6935
HUMAN NECESSITIES
A61P35/00
HUMAN NECESSITIES
International classification
A61K47/69
HUMAN NECESSITIES
A61K31/4745
HUMAN NECESSITIES
C08G69/48
CHEMISTRY; METALLURGY
A61K31/704
HUMAN NECESSITIES
Abstract
Described herein are platinum-based brush-arm star polymers (Pt-BASPs), or a pharmaceutical composition thereof, for delivery of platinum-based agents, such as oxaliplatin. Also provided are methods and kits involving the Pt-BASPs, or a pharmaceutical composition thereof, for treating proliferative diseases such as cancers (e.g., lung cancer, head-and-neck cancer, esophagus cancer, stomach cancer, breast cancer, pancreas cancer, colorectal cancer, liver cancer, kidney cancer, or prostate cancer) in a subject.
Claims
1. A platinum complex of Formula (I): ##STR00052## or a salt thereof, wherein: each instance of R.sup.N1 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group, or two R.sup.N1 are taken with the intervening atoms to form a heterocyclic ring; each instance of R.sup.N2 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group, or two R.sup.N2 are taken with the intervening atoms to form a heterocyclic ring; L.sup.N is a linker selected from the group consisting of substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heterarylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocyclylene, and combinations thereof; and each instance of n is 1, 2, 3, 4, 5, or 6.
2. The platinum complex of claim 1, wherein the platinum complex is of Formula (I-a): ##STR00053## or a salt thereof, wherein: each instance of R.sup.A is independently hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; and each instance of R.sup.B is independently hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; or two instances of R.sup.B are joined to form a substituted or unsubstituted, carbocyclic ring, or substituted or unsubstituted, heterocyclic ring.
3. The platinum complex of claim 2, wherein the platinum complex is of Formula (I-b): ##STR00054## or a salt thereof.
4. The platinum complex of claim 2, wherein the platinum complex is of Formula (I-c): ##STR00055## or a salt thereof, wherein: Ring A is a substituted or unsubstituted, monocyclic, 3- to 7-membered carbocyclic ring; each instance of R.sup.C is independently halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR.sup.a, —N(R.sup.a).sub.2, —SR.sup.a, —CN, —SCN, —C(═NR.sup.a)R.sup.a, —C(═NR.sup.a)OR.sup.a, —C(═NR.sup.a)N(R.sup.a).sub.2, —C(═O)R.sup.a, —C(═O)OR.sup.a, —C(═O)N(R.sup.a).sub.2, —NO.sub.2, —N(R.sup.a)C(═O)R.sup.a, —N(R.sup.a)C(═O)OR.sup.a, —N(R.sup.a)C(═O)N(R.sup.a).sub.2, —N(R.sup.a)S(═O)R.sup.a, —N(R.sup.a)S(═O)OR.sup.a, —N(R.sup.a)S(═O)N(R.sup.a).sub.2, —N(R.sup.a)S(═O).sub.2R.sup.a, —N(R.sup.a)S(═O).sub.2OR.sup.a, —N(R.sup.a)S(═O).sub.2N(R.sup.a).sub.2, —OC(═O)R.sup.a, —OC(═O)OR.sup.a, or —OC(═O)N(R.sup.a).sub.2; each instance of R.sup.a is independently hydrogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, or a sulfur protecting group when attached to a sulfur atom, or two instances of R.sup.a are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring; and u is an integer between 0 and 8, inclusive.
5. The platinum complex of claim 4, wherein the platinum complex is of Formula (I-d): ##STR00056## or a salt thereof.
6. The platinum complex of claim 4, wherein the platinum complex is of Formula (I-e): ##STR00057## or a salt thereof, wherein u is an integer between 0 and 8, inclusive.
7. The platinum complex of claim 1, wherein the platinum complex is of Formula (I-f): ##STR00058## or a salt thereof.
8. (canceled)
9. The platinum complex of claim 1, or a salt thereof, wherein each instance of R.sup.N1 is hydrogen, and each instance of R.sup.N2 is hydrogen.
10-11. (canceled)
12. The platinum complex of claim 1, or a salt thereof, wherein L.sup.N is substituted or unsubstituted heterocyclylene or substituted or unsubstituted cycloalkylene.
13-14. (canceled)
15. The platinum complex of claim 1, or a salt thereof, wherein each instance of ##STR00059## is of the formula: ##STR00060##
16-18. (canceled)
19. The platinum complex of claim 1, wherein the platinum complex is of the formula: ##STR00061##
20. The platinum complex of claim 1, wherein the platinum complex is of the formula: ##STR00062##
21. A method of preparing a platinum complex of claim 1, or a salt thereof, the method comprising the steps of: (a) oxidizing a compound of Formula (S-1): ##STR00063## with an oxidant to provide a compound of Formula (S-2): ##STR00064## and (b) coupling the compound of Formula (S-2) with a compound of Formula (S-3): ##STR00065## to provide the platinum complex, or the salt thereof.
22-24. (canceled)
25. A polymer prepared by the steps of: (a) reacting a first macromonomer of Formula (III′): ##STR00066## or a salt thereof, with a second macromonomer of Formula (III′), or a salt thereof, in the presence of a metathesis catalyst to provide a polymerization mixture, wherein: a is an integer from 1 to 10, inclusive; each instance of b is independently an integer from 1 to 10, inclusive; c is an integer from 1 to 200, inclusive; e is 0, 1, 2, 3, 4, 5, or 6; each instance of L is independently —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, a polymer, or a substituted or unsubstituted C.sub.1-30 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is independently replaced with substituted or unsubstituted phenyl, substituted or unsubstituted triazolyl, —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, substituted or unsubstituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, or two R.sup.Lb groups are joined to form a substituted or unsubstituted carbocyclic or substituted or unsubstituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form a substituted or unsubstituted heterocyclic ring; each instance of M is independently hydrogen or an agent; and the first macromonomer and the second macromonomer are the same or different from each other; and (b) contacting the polymerization mixture with a platinum complex of claim 1, or a salt thereof, to provide the polymer.
26-27. (canceled)
28. A polymer prepared by the steps of: (a) reacting a first macromonomer of Formula (III): ##STR00067## or a salt thereof, with a second macromonomer of Formula (III), or a salt thereof, in the presence of a metathesis catalyst to provide a polymerization mixture, wherein: a is an integer from 1 to 10, inclusive; each instance of b is independently an integer from 1 to 5, inclusive; c is an integer from 1 to 100, inclusive; e is 0, 1, 2, 3, or 4; each instance of L is independently —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, or a substituted or unsubstituted C.sub.1-10 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is replaced with —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O).sup.−, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, substituted or unsubstituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, or two R.sup.Lb groups are joined to form a substituted or unsubstituted carbocyclic or substituted or unsubstituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form a substituted or unsubstituted heterocyclic ring; each instance of M is independently hydrogen or a pharmaceutical agent; and the first macromonomer and the second macromonomer are the same or different from each other; and (b) contacting the polymerization mixture with a platinum complex of claim 1, or a salt thereof, to provide the polymer.
29-48. (canceled)
49. A pharmaceutical composition comprising a polymer of claim 25 and optionally a pharmaceutically acceptable excipient.
50. A kit comprising: a polymer of claim 25; and instructions for using the polymer.
51. A method of treating a proliferative disease in a subject in need thereof comprising administering to the subject an effective amount of a pharmaceutical composition of claim 49.
52-54. (canceled)
55. A method of delivering a pharmaceutical agent to a subject comprising administering to the subject a pharmaceutical composition of claim 49.
56. (canceled)
57. A method of delivering a pharmaceutical agent to a biological sample, tissue, or cell, the method comprising contacting the biological sample, tissue, or cell with a pharmaceutical composition of claim 49.
58. (canceled)
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0105] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.
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DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION
[0135] Oxaliplatin and related derivatives are widely used in cancer chemotherapy. Significant adverse reactions related to oxaliplatin and related derivatives frequently limits the use of higher doses to achieve their maximum antineoplastic effects. The present invention provides platinum-based brush-arm star polymers (Pt-BASPs) and methods for the controlled delivery of oxaliplatin and related compounds. In certain embodiments, the Pt-BASPs are loaded with a bis-norbornene oxaliplatin crosslinker (oxPt-XL). In certain embodiments, the Pt-BASPs are loaded with oxPt-XL and one or more therapeutic, diagnostic, or prophylactic agents for multi-agent delivery.
Oxaliplatin or Its Derivative Crosslinkers
[0136] In one aspect, the present invention provides a platinum complex of Formula (I):
##STR00007##
or salts thereof,
wherein:
[0137] each instance of R.sup.N1 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, a nitrogen protecting group, or two R.sup.N1 are taken with the intervening atoms to form a heterocyclic ring;
[0138] each instance of R.sup.N2 is independently hydrogen, substituted or unsubstituted C.sub.1-6 alkyl, or a nitrogen protecting group, or or two R.sup.N2 are taken with the intervening atoms to form a heterocyclic ring;
[0139] L.sup.N is a linker selected from the group consisting of substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heterarylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocyclylene, and combinations thereof; and
[0140] each instance of n is 1, 2, 3, 4, 5, or 6.
[0141] As generally defined herein, each instance of R.sup.N1 is independently hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting group, or two R.sup.N1 are taken with the intervening atoms to form a heterocyclic ring. In certain embodiments, at least one instance of R.sup.N1 is hydrogen. In certain embodiments, each instance of R.sup.N1 is hydrogen. In certain embodiments, at least one instance of R.sup.N1 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, each instance of R.sup.N1 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.N1 is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.N1 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.N1 is methyl. In certain embodiments, R.sup.N1 is ethyl. In certain embodiments, R.sup.N1 is a nitrogen protecting group. In certain embodiments, two R.sup.N1 are taken with the intervening atoms to form a heterocyclic ring.
[0142] As generally defined herein, each instance of R.sup.N2 is independently hydrogen, optionally substituted C.sub.1-6 alkyl, or a nitrogen protecting group, or two R.sup.N2 are taken with the intervening atoms to form a heterocyclic ring, or R.sup.N1 and R.sup.N2 are taken with the intervening atoms to form a heterocyclic ring. In certain embodiments, at least one instance of R.sup.N2 is hydrogen. In certain embodiments, each instance of R.sup.N2 is hydrogen. In certain embodiments, at least one instance of R.sup.N2 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, each instance of R.sup.N2 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.N2 is substituted C.sub.1 6 alkyl. In certain embodiments, R.sup.N2 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.N2 is methyl. In certain embodiments, R.sup.N2 is ethyl. In certain embodiments, R.sup.N2 is a nitrogen protecting group. In certain embodiments, two R.sup.N2 are taken with the intervening atoms to form a heterocyclic ring. In certain embodiments, R.sup.N1 and R.sup.N2 are taken with the intervening atoms to form a heterocyclic ring.
[0143] In certain embodiments, R.sup.N1 and R.sup.N2 are the same. In certain embodiments, R.sup.N1 and R.sup.N2 are different. In certain embodiments, all instances of R.sup.N1 and R.sup.N2 are hydrogen. In certain embodiments, at least one instance of R.sup.N1 is hydrogen and at least one instance of R.sup.N2 is not hydrogen. In certain embodiments, at least one instance of R.sup.N1 is hydrogen and at least one instance of R.sup.N2 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, at least one instance of R.sup.N1 is hydrogen and at least one instance of R.sup.N2 is substituted C.sub.1-6 alkyl. In certain embodiments, at least one instance of R.sup.N1 is hydrogen and at least one instance of R.sup.N2 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, at least one instance of R.sup.N1 is hydrogen and at least one instance of R.sup.N2 is methyl, ethyl, or propyl. In certain embodiments, at least one instance of R.sup.N1 and at least one instance of R.sup.N2 are each independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, at least one instance of R.sup.N1 and at least one instance of R.sup.N2 are each independently substituted C.sub.1-6 alkyl. In certain embodiments, at least one instance of R.sup.N1 and at least one instance of R.sup.N2 are each independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, at least one instance of R.sup.N1 and at least one instance of R.sup.N2 are each independently is methyl, ethyl, or propyl. In certain embodiments, at least one instance of R.sup.N1 and at least one instance of R.sup.N2 are each independently a nitrogen protecting group.
[0144] As generally defined herein, L.sup.N is a linker selected from the group consisting of substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heterarylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocyclylene, and combinations (e.g., combination of two, three, or four) thereof. In certain embodiments, L.sup.N is substituted or unsubstituted alkylene. In certain embodiments, L.sup.N is substituted alkylene. In certain embodiments, L.sup.N is unsubstituted alkylene (e.g., methylene, ethylene, or propylene). In certain embodiments, L.sup.N is substituted or unsubstituted heteroalkylene. In certain embodiments, L.sup.N is substituted or unsubstituted arylene. In certain embodiments, L.sup.N is substituted or unsubstituted phenylene. In certain embodiments, L.sup.N is substituted or unsubstituted heterarylene. In certain embodiments, L.sup.N is substituted or unsubstituted 5-membered or 6-membered heterarylene. In certain embodiments, L.sup.N is substituted or unsubstituted cycloalkylene. In certain embodiments, L.sup.N is substituted or unsubstituted cyclohexylene. In certain embodiments, L.sup.N is substituted or unsubstituted cyclohexylene, wherein the point of attachments are 1- and 4-positions. In certain embodiments, L.sup.N is substituted or unsubstituted cyclohexylene, wherein the point of attachments are 1- and 3-positions. In certain embodiments, L.sup.N is substituted or unsubstituted cyclohexylene, wherein the point of attachments are 1- and 2-positions.
[0145] In certain embodiments, L.sup.N is substituted or unsubstituted cycloalkylene and R.sup.N1 and R.sup.N2 are the same. In certain embodiments, L.sup.N is substituted or unsubstituted cyclohexylene and R.sup.N1 and R.sup.N2 are hydrogen. In certain embodiments, L.sup.N is substituted or unsubstituted cyclohexylene, wherein the point of attachments are 1- and 4-positions, and R.sup.N1 and R.sup.N2 are hydrogen.
[0146] In certain embodiments, the number of atoms in the shortest backbone of L.sup.N is 1. In certain embodiments, the number of atoms in the shortest backbone of L.sup.N is 2. In certain embodiments, the number of atoms in the shortest backbone of L.sup.N is 3. In certain embodiments, the number of atoms in the shortest backbone of L.sup.N is 4.
[0147] As generally defined herein, n is 1, 2, 3, 4, 5, or 6. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5. In certain embodiments, n is 6.
[0148] In certain embodiments, the platinum complexes of Formula (I) are prodrugs of the oxaliplatin. The Pt—O bond can be cleaved by a reducing agent in vivo to release oxaliplatin. In certain embodiments, the platinum complex of Formula (I) is used as a crosslinker to prepare polymers for the controlled release of oxaliplatin. In certain embodiments, the platinum complex of Formula (I) is used as a crosslinker to prepare Pt-BASP.
[0149] In certain embodiments, the platinum complex of Formula (I) is of Formula (I-a):
##STR00008##
or a salt thereof,
wherein:
[0150] each instance of R.sup.A is independently hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; and
[0151] each instance of R.sup.B is independently hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
[0152] or two instances of R.sup.B are joined to form a substituted or unsubstituted, carbocyclic ring, or substituted or unsubstituted, heterocyclic ring.
[0153] In certain embodiments, the platinum complex of Formula (I) is of Formula (I-b):
##STR00009##
or a salt thereof.
[0154] In certain embodiments, at least one instance of R.sup.A is hydrogen. In certain embodiments, each instance of R.sup.A is hydrogen. In certain embodiments, at least one instance of R.sup.A is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In certain embodiments, at least one instance of R.sup.A is unsubstituted alkyl (e.g., unsubstituted C.sub.1-6 alkyl, such as methyl, ethyl, or propyl). In certain embodiments, at least one instance of R.sup.A is substituted alkyl (e.g., substituted C.sub.1-6 alkyl). In certain embodiments, at least one instance of R.sup.A is halogen (e.g., F).
[0155] In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.A is hydrogen. In certain embodiments of Formulae (I-a)-(I-b), both instances of R.sup.A are hydrogen. In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.A is substituted or unsubstituted alkyl. In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.A is unsubstituted alkyl. In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.A is substituted alkyl.
[0156] In certain embodiments, at least one instance of R.sup.B is hydrogen. In certain embodiments, each instance of R.sup.B is hydrogen. In certain embodiments, at least one instance of R.sup.B is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In certain embodiments, at least one instance of R.sup.B is unsubstituted alkyl (e.g., unsubstituted C.sub.1-6 alkyl, such as methyl, ethyl, or propyl). In certain embodiments, at least one instance of R.sup.B is substituted alkyl (e.g., substituted C.sub.1-6 alkyl). In certain embodiments, at least one instance of R.sup.B is halogen (e.g., F).
[0157] In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.B is hydrogen. In certain embodiments of Formulae (I-a)-(I-b), both instances of R.sup.B are hydrogen. In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.B is substituted or unsubstituted alkyl. In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.B is unsubstituted alkyl. In certain embodiments of Formulae (I-a)-(I-b), at least one instance of R.sup.B is substituted alkyl. In certain embodiments of Formulae (I-a)-(I-b), two R.sup.B are joined to form a substituted or unsubstituted, carbocyclic ring, or substituted or unsubstituted, heterocyclic ring. In certain embodiments of Formulae (I-a)-(I-b), two R.sup.B are joined to form a substituted or unsubstituted carbocyclic ring. In certain embodiments of Formulae (I-a)-(I-b), two R.sup.B are joined to form a substituted or unsubstituted cyclohexyl ring.
[0158] In certain embodiments, the platinum complex of Formula (I) is of Formula (I-c):
##STR00010##
or a salt thereof,
wherein:
[0159] Ring A is a substituted or unsubstituted, monocyclic, 3- to 7-membered carbocyclic ring;
[0160] each instance of R.sup.C is independently halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR.sup.a, —N(R.sup.a).sub.2, —SR.sup.a, —CN, —SCN, —C(═NR.sup.a)R.sup.a, —C(═NR.sup.a)OR.sup.a, —C(═NR.sup.a)N(R.sup.a).sub.2, —C(═O)R.sup.a, —C(═O)OR.sup.a, —C(═O)N(R.sup.a).sub.2, —N.sub.02, —N(R.sup.a)C(═O)R.sup.a, —N(R.sup.a)C(═O)OR.sup.a, —N(R.sup.a)C(═O)N(R.sup.a).sub.2, —N(R.sup.a)S(═O)R.sup.a, —N(R.sup.a)S(═O)OR.sup.a, —N(R.sup.a)S(═O)N(R.sup.a).sub.2, —N(R.sup.a)S(═O).sub.2R.sup.a, —N(R.sup.a)S(═O).sub.2OR.sup.a, —N(R.sup.a)S(═O).sub.2N(R.sup.a).sub.2, —OC(═O)R.sup.a, —OC(═O)OR.sup.a, or —OC(═O)N(R.sup.a).sub.2;
[0161] each instance of R.sup.a is independently hydrogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, or a sulfur protecting group when attached to a sulfur atom, or two instances of R.sup.a are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring; and
[0162] u is an integer between 0 and 8, inclusive.
[0163] In certain embodiments, the platinum complex of Formula (I) is of Formula (I-d):
##STR00011##
or a salt thereof.
[0164] In certain embodiments of Formulae (I-c) and (I-d), Ring A is a substituted or unsubstituted cyclopropyl ring. In certain embodiments of Formulae (I-c) and (I-d), Ring A is a substituted or unsubstituted cyclobutyl ring. In certain embodiments of Formulae (I-c) and (I-d), Ring A is a substituted or unsubstituted cyclopentyl ring. In certain embodiments of Formulae (I-c) and (I-d), Ring A is a substituted or unsubstituted cyclohexyl ring. In certain embodiments of Formulae (I-c) and (I-d), Ring A is a substituted or unsubstituted cycloheptyl ring.
[0165] In certain embodiments of Formula (I-d), R.sup.A is hydrogen. In certain embodiments of Formula (I-d), R.sup.A is substituted or unsubstituted alkyl. In certain embodiments of Formula (I-d), R.sup.A is unsubstituted alkyl (e.g., methyl, ethyl, or propyl). In certain embodiments of Formula (I-d), R.sup.A is substituted alkyl.
[0166] In certain embodiments, at least one instance of R.sup.C is hydrogen. In certain embodiments, each instance of R.sup.C is hydrogen. In certain embodiments, at least one instance of R.sup.C is substituted or unsubstituted alkyl (e.g., substituted or unsubstituted C.sub.1-6 alkyl). In certain embodiments, at least one instance of R.sup.C is unsubstituted alkyl (e.g., unsubstituted C.sub.1-6 alkyl, such as methyl, ethyl, or propyl). In certain embodiments, at least one instance of R.sup.C is substituted alkyl (e.g., substituted C.sub.1-6 alkyl). In certain embodiments, at least one instance of R.sup.C is halogen (e.g., F). In certain embodiments, each instance of R.sup.C is the same.
[0167] In certain embodiments, the platinum complex of Formula (I) is of Formula (I-e):
##STR00012##
or a salt thereof.
[0168] In certain embodiments of Formulae (I-c)-(I-e), u is 0. In certain embodiments of Formulae (I-c)-(I-e), u is 2. In certain embodiments of Formulae (I-c)-(I-e), u is 3. In certain embodiments of Formulae (I-c)-(I-e), u is 4. In certain embodiments of Formulae (I-c)-(I-e), u is 5. In certain embodiments of Formulae (I-c)-(I-e), u is 6. In certain embodiments of Formulae (I-c)-(I-e), u is 7. In certain embodiments of Formulae (I-c)-(I-e), u is 8.
[0169] In certain embodiments, the platinum complex of Formula (I) is of Formula (I-f):
##STR00013##
or a salt thereof.
[0170] As generally used herein,
##STR00014##
encompasses a racemic moiety or any stereoisomer thereof. In certain embodiments, at least one instance of
##STR00015##
is of one of the following formulae:
##STR00016##
In certain embodiments, each instance of
##STR00017##
is of one of the following formulae:
##STR00018##
In certain embodiments, each instance of
##STR00019##
is of
the formula:
##STR00020##
[0171] In certain embodiments, the provided platinum complex is of the formula:
##STR00021##
[0172] In certain embodiments, the provided platinum complex is of the formula:
##STR00022##
[0173] The platinum complex of Formula (I) can be prepared using the general methodology shown in Scheme 1. Details of the synthetic procedures are described in the Examples below.
##STR00023##
[0174] In one embodiments, the invention provides methods of preparing a platinum complex of Formula (I), the method comprising steps of:
[0175] (a) oxidizing a compound of Formula (S-1) with an oxidant
##STR00024##
[0176] to yield a compound of Formula (S-2):
##STR00025##
wherein R.sup.N1, R.sup.N2, L.sup.N, and n are as defined herein; and
[0177] (b) coupling the compound of Formula (S-2) with a compound of Formula (S-3):
##STR00026##
to yield a platinum complex of Formula (I).
[0178] In certain embodiments, the oxidant used in step (a) can oxidize Pt (II) to Pt (IV) with two hydroxyl groups under suitable oxidization condition (Hall et al., J. Biol. Inorg. Chem. 2003, 8, 726). In certain embodiments, the oxidant is H.sub.2O.sub.2.
[0179] In certain embodiments, an activator is present in the coupling reaction in step (b). The activator converts the compound of Formula (S-3) to an activated ester for the coupling reaction. Examples of useful activators are dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (WSC/HCl), diphenylphosphorylazide (DPPA), carbonyldiimidazole (CDI), diethylcyanophosphonate (DEPC), benzotriazole-1-yloxy-trispyrrolidinophosphonium (DIPCI), benzotriazole-1-yloxy-trispyrrolidinophosphonium hexafluorophosphate (PyBOP), 1-hydroxybenzotriazole (HOBt), hydroxysuccinimide (HOSu), dimethylaminopyridine (DMAP), 1-hydroxy-7-azabenzotriazole (HOAt), hydroxyphthalimide (HOPht), pentafluorophenol (Pfp-OH), 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphonate (HATU), O-benzotriazole-1-yl-1,1,3,3-tetramethyluronium tetrafluoroborate (TBTU), 3,4-dihydro-3-hydrodi-4-oxa-1,2,3-benzotriazine (Dhbt). In certain embodiments, the activator is DCC.
[0180] The activator is used in an amount of about 1 to 20 equivalents of the compound of Formula (S-2). In certain embodiments, the activator is used in an amount of about 1 to 10 equivalents. In certain embodiments, the activator is used in an amount of about 1 to 5 equivalents.
[0181] Examples of useful solvents in the coupling reaction are DMSO, DMF, methylene chloride. The coupling reaction can be conducted at 0 to 50° C. In certain embodiments, the coupling reaction is conducted at room temperature for about 10 min to about 30 hours. In certain embodiments, the coupling reaction is conducted for about 15 minutes to about 24 hours.
Platinum-based Brush-Arm Star Polymers (Pt-BASPs)
[0182] The present invention provides platinum-based brush-arm star polymers (Pt-BASP) using the “brush-first” ring-opening metathesis polymerization (ROMP). The brush-first method involves sequential copolymerization of two functional monomers, a polymeric macromonomer (MM) followed by a multifunctional crosslinker, to generate a unimolecular micelle-like nanostructure with a core comprised of oxaliplatin or a derivative thereof crosslinker and a corona comprised of the MM. Synthesis of Pt-BASPs is generally described in International Application No. PCT/US2014/033554 filed Apr. 9, 2014, which is incorporated by reference in its entirety herein.
[0183] A Pt-BASP described herein includes oxaliplatin or a derivative thereof attached to the rest of the Pt-BASP through ester bonds. Oxaliplatin is a clinically approved chemotherapeutic agent that includes a platinum(II) core, the bidentate ligand, 1,2-diaminocyclohexane, and a bidentate oxalate group.
[0184] In addition to oxaliplatin or a derivative thereof, a Pt-BASP described herein may include one or more additional agents that are not oxaliplatin or a derivative thereof to form multi-agent-loaded (e.g., multi-drug-loaded) Pt-BASPs. The described Pt-BASPs are advantageous over known nanoparticle (NP)-based delivery systems. NP-based combination cancer therapy has the potential to overcome the toxicity and poorly controlled dosing of traditional systemic combination therapies (Hu, C. M. J.; Zhang, L. F. Biochem. Pharmacol. 2012, 83, 1104; Yan, Y.; Bjornmalm, M.; Caruso, F. ACS Nano 2013, 7, 9512; Ma, L.; Kohli, M.; Smith, A. ACS Nano 2013, 7, 9518). Though NP-based therapeutics for cancer therapy have been the subject of numerous investigations over the past several decades (Duncan, R. Nat. Rev. Drug Discovery 2003, 2, 347; Peer et al. Nat. Nanotechnol. 2007, 2, 751; Wolinsky, J. B.; Grinstaff, M. W. Adv. Drug Delivery Rev. 2008, 60, 1037; Davis, M. E.; Chen, Z.; Shin, D. M. Nat. Rev. Drug Discovery 2008, 7, 771; Kwon, G. S.; Kataoka, K. Adv. Drug Delivery Rev. 2012, 64, 237), ratiometric, synchronized release of multiple drugs from single NP scaffolds remains a challenge (Sengupta et al. Nature 2005, 436, 568; Lammers et al. Biomaterials 2009, 30, 3466; Kolishetti, N.; Dhar, S.; Valencia, P. M.; Lin, L. Q.; Karnik, R.; Lippard, S. J.; Langer, R.; Farokhzad, O. C. Proc. Natl. Acad. Sci. U.S.A 2010, 107, 17939; Aryal, S.; Hu, C. M. J.; Zhang, L. F. Mol. Pharm. 2011, 8, 1401). Many reported nanoparticle architectures for delivery, e.g., liposomes, micelles, and dendrimers, are not readily amenable to controlled incorporation and release of multiple drugs.
[0185] In contrast, the Pt-BASPs described herein are able to deliver multiple agents (e.g., cisplatin and one or more other agents) ratiometrically. The agents included in a Pt-BASP may show different therapeutic, diagnostic, and/or prophylactic effects on a subject, tissue, or cell. For example, a Pt-BASP may include two or more therapeutic agents (including oxaliplatin or a derivative thereof), and the therapeutic agents may show different anti-proliferative activities (e.g., anti-cancer activities) at each therapeutic agent's maximum tolerated dose (MTD). A key benefit of single nanoparticle combination therapy is the ability to deliver multiple drugs at a precise ratio to a subject, tissue, or cell, while minimizing undesired effects (e.g., toxicity) associated with multiple drug combinations. To achieve the maximum therapeutic index in a multi-drug combination therapy, simultaneous dosing of each drug at or near each drug's MTD would be ideal. A Pt-BASP described herein may include multiple drugs at or near each drug's MTD before the Pt-BASP is delivered to a subject, tissue, or cell, release the multiple drugs at or near each drug's MTD into the subject, tissue, or cell after delivery, and therefore achieve the maximum therapeutic index. In certain embodiments, Pt-BASPs described herein include camptothecin (CPT) and oxaliplatin or a derivative thereof (e.g., oxPt-XL). In certain embodiments, Pt-BASPs described herein include irinotecan (IRT) and oxaliplatin or a derivative thereof (e.g., oxPt-XL). In certain embodiments, Pt-BASP described herein include SN-38 and oxaliplatin or a derivative thereof (e.g. oxPt-XL). In certain embodiments, Pt-BASPs described herein include 5-FU and oxaliplatin or a derivative thereof (e.g., oxPt-XL). In certain embodiments, Pt-BASPs described herein include IRT, 5-FU, and oxaliplatin or a derivative thereof. In certain embodiments, Pt-BASPs described herein include CPT, 5-FU, and oxaliplatin or a derivative thereof. In certain embodiments, Pt-BASPs described herein include SN-38, 5-FU, and oxaliplatin or a derivative thereof.
[0186] The Pt-BASPs described herein are also able to deliver multiple agents orthogonally. Different chemical and/or physical conditions may be employed to individually release the multiple agents upon delivery. The convergent synthesis of Pt-BASPs allow the attachment of different agents to the Pt-BASPs through different linkers (e.g., linkers cleavable by reduction, such as Pt—O bonds; hydrolysable linkers, such as ester bonds; and photo-cleavable linkers, such
as the moiety
##STR00027##
wherein the moiety may be further substituted). For example, oxaliplatin may be released from a Pt-BASP by a reduction reaction of the Pt—O bonds; and other agents included in the Pt-BASP may be released from the Pt-BASP under chemical and/or physical conditions that are different from the reduction reaction. In certain embodiments, an agent, other than oxaliplatin or a derivative thereof, included in a Pt-BASP is released from the Pt-BASP by hydrolysis (e.g., hydrolysis under acidic conditions). In certain embodiments, an agent, other than oxaliplatin or a derivative thereof, included in a Pt-BASP is released from the Pt-BASP by irradiation with ultraviolet light (UV).
[0187] The Pt-BASPs described herein can be directly constructed using carefully designed drug-conjugates as building blocks, and no self-assembly is required for preparing the Pt-BASPs. The methods for preparing the Pt-BASPs described herein involves ring-opening metathesis polymerization (ROMP) (Liu et al. J. Am. Chem. Soc. 2012, 134, 16337; Liu, J.; Gao, A. X.; Johnson, J. A. J Vis Exp 2013, e50874). In certain embodiments, the Pt-BASPs described herein are prepared by polymerization of norbornene-terminated macromonomers (MMs) followed by in situ crosslinking with bis-norbornene crosslinkers. The preparation methods described herein are versatile and have little limitations, e.g., in terms of the different agents that can be built into the Pt-BASPs. In certain embodiments, an agent that can be built into the Pt-BASPs includes addressable functional groups that are compatible with ROMP. In certain embodiments, the invention provides Pt-BASPs prepared by Method A′ including the steps of:
[0188] (a) reacting a macromonomer of Formula (III′):
##STR00028##
or a salt thereof, with a metathesis catalyst to form a polymerization mixture, wherein: [0189] a is an integer from 1 to 10, inclusive; [0190] each instance of b is independently an integer from 1 to 10, inclusive; [0191] c is an integer from 1 to 200, inclusive; [0192] e is 0, 1, 2, 3, 4, 5, or 6; [0193] each instance of L is independently —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, a polymer, or a substituted or unsubstituted C.sub.1-30 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is independently replaced with substituted or unsubstituted phenyl, substituted or unsubstituted triazolyl, —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, substituted or unsubstituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, or two R.sup.Lb groups are joined to form a substituted or unsubstituted carbocyclic or substituted or unsubstituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form a substituted or unsubstituted heterocyclic ring;
[0194] each instance of M is independently hydrogen or an agent (e.g., pharmaceutical agent (e.g., therapeutic, diagnostic, or prophylactic agent)); and
[0195] (b) contacting the polymerization mixture from step (a) with a platinum complex of Formula (I) (e.g., a solution of a platinum complex of Formula (I)).
[0196] In certain embodiments, the invention provides Pt-BASPs prepared by Method A including the steps of:
[0197] (a) reacting a macromonomer of Formula (III)
##STR00029##
with a metathesis catalyst to form a polymerization mixture;
wherein:
[0198] a is an integer from 1 to 10, inclusive;
[0199] each instance of b is independently an integer from 1 to 5 inclusive;
[0200] c is an integer from 30 to 100 inclusive;
[0201] e is 0, 1, 2, 3, or 4;
[0202] each instance of L is independently —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, or an optionally substituted C.sub.1-10 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is replaced with —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, optionally substituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, or two R.sup.Lb groups are joined to form an optionally substituted carbocyclic or optionally substituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form an optionally substituted heterocyclic ring; and
[0203] each instance of M is independently hydrogen or a pharmaceutical agent (e.g., therapeutic, diagnostic, or prophylactic agent); and
[0204] (b) contacting the polymerization mixture from step (a) with a platinum complex of Formula (I) (e.g., a solution of a platinum complex of Formula (I)).
[0205] In certain embodiments, M in the macromonomer is oxaliplatin or a derivative thereof. In certain embodiments, M in the macromonomer is oxPt-XL. In Method A′ (e.g., Method A), step (a) may be performed in the presence of a non-agent-loaded MM, such as a macromonomer of Formula (IV):
##STR00030##
In certain embodiments, the molar ratio of a non-agent-loaded MM to the combined agents (including oxPt-XL) is about 0.01:1, about 0.3:1, about 0.67:1, about 1:1, about 1.5:1, about 3:1, about 10:1, about 30:1, or about 100:1, inclusive. In certain embodiments, the molar ratio of a non-agent-loaded macromonomer to the combined agents (including cisplatin) is about 0.67:1, inclusive.
[0206] In certain embodiments, the provided Pt-BASPs are prepared by Method B′ including the steps of:
[0207] (a) reacting a first macromonomer of Formula (III′), or a salt thereof, with a second macromonomer of Formula (III′), or a salt thereof:
##STR00031##
in the presence of a metathesis catalyst to form a polymerization mixture, wherein a, L, M, b, e, and c are as described herein, provided that M in the first macromonomer is different from M in the second macromonomer; and
[0208] (b) contacting the polymerization mixture from step (a) with a platinum complex of Formula (I) (e.g., a solution of a platinum complex of Formula (I)).
[0209] In certain embodiments, the provided Pt-BASPs are loaded with more than one therapeutic, diagnostic, or prophylactic agents other than oxaliplatin or a derivative thereof and can be prepared by Method B including the steps of:
[0210] (a) reacting a first macromonomer of Formula (III) with a second macromonomer of Formula (III)
##STR00032##
in the presence of a metathesis catalyst to form a polymerization mixture,
wherein:
[0211] a is an integer from 1 to 10, inclusive;
[0212] each instance of b is independently an integer from 1 to 5 inclusive;
[0213] c is an integer from 1 to 100 inclusive;
[0214] e is 0, 1, 2, 3, or 4;
[0215] each instance of L is independently —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, or an optionally substituted C.sub.1-10 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is replaced with —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, optionally substituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, or two R.sup.Lb groups are joined to form an optionally substituted carbocyclic or optionally substituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form an optionally substituted heterocyclic ring;
[0216] each instance of M is independently hydrogen or a pharmaceutical agent (e.g., therapeutic, diagnostic, or prophylactic agent); and
[0217] M in the first macromonomer is different from M in the second macromonomer; and
[0218] (b) contacting the polymerization mixture from step (a) with a platinum complex of Formula (I) (e.g., a solution of a platinum complex of Formula (I)).
[0219] In Method B′ (e.g., Method B), step (a) may be performed in the presence of a non-agent-loaded MM, such as a macromonomer of Formula (IV):
##STR00033##
In certain embodiments, the non-agent-loaded MM is macromonomer PEG-MM. In certain embodiments of the Pt-BASPs loaded with more than one therapeutic, diagnostic, or prophylactic agents, Ms in the first and second macromonomers are different. In certain embodiments of the Pt-BASPs loaded with more than one therapeutic, diagnostic, or prophylactic agents, Ms in the first and second macromonomers are the same. In certain embodiments of the Pt-BASPs loaded with more than one therapeutic, diagnostic, or prophylactic agents, at one instance of Ms in the first MM and at least one instance of Ms in the second MM are different from each other. In certain embodiments, Ms in the first and second macromonomers are not oxaliplatin or a derivative thereof (e.g., oxPt-XL). In certain embodiments of the Pt-BASPs loaded with more than one therapeutic, diagnostic, or prophylactic agents, Ms in the first and second macromonomers are different therapeutic agents. In certain embodiments, M in the first macromonomer is camptothecin, and M in the second macromonomer is 5-FU. In certain embodiments, M in the first macromonomer is IRT, and M in the second macromonomer is 5-FU. In certain embodiments, M in the first macromonomer is SN-38, and M in the second macromonomer is 5-FU.
[0220] As generally defined herein, a is an integer from 1 to 10, inclusive. In certain embodiments, a is 1. In certain embodiments, a is 2. In certain embodiments, a is 3. In certain embodiments, a is 4. In certain embodiments, a is 5. In certain embodiments, a is 6. In certain embodiments, a is 7. In certain embodiments, a is 8. In certain embodiments, a is 9. In certain embodiments, a is 10.
[0221] As generally defined herein, each instance of b is independently an integer from 1 to 10, inclusive. In certain embodiments, b is an integer from 1 to 5, inclusive. In certain embodiments, b is 1. In certain embodiments, b is 2. In certain embodiments, b is 3. In certain embodiments, b is 4. In certain embodiments, b is 5. In certain embodiments, b is 6. In certain embodiments, b is 7. In certain embodiments, b is 8. In certain embodiments, b is 9. In certain embodiments, b is 10.
[0222] As generally defined herein, c is an integer from 1 to 200, inclusive. In certain embodiments, c is an integer from 1 to 100, inclusive. In certain embodiments, c is an integer from 1 to 29, inclusive. In certain embodiments, c is an integer from 30 to 100, inclusive. In certain embodiments, c is an integer from 40 to 90, inclusive. In certain embodiments, c is an integer from 50 to 80, inclusive. In certain embodiments, c is an integer from 60 to 70, inclusive. In certain embodiments, c is about 65. In certain embodiments, c is about 66. In certain embodiments, c is about 67. In certain embodiments, c is about 68. In certain embodiments, c is about 69. In certain embodiments, c is about 70. In certain embodiments, c is an integer from 101 to 200, inclusive.
[0223] As generally defined herein, e is 0, 1, 2, 3, 4, 5, or 6. In certain embodiments, e is 0, 1, 2, 3, or 4. In certain embodiments, e is 0. In certain embodiments, e is 1. In certain embodiments, e is 2. In certain embodiments, e is 3. In certain embodiments, e is 4. In certain embodiments, e is 5. In certain embodiments, e is 6.
[0224] As generally defined herein, each instance of L is independently —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, a polymer, or a substituted or unsubstituted C.sub.1-30 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is independently replaced with substituted or unsubstituted phenyl, substituted or unsubstituted triazolyl, —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C.sup.−, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, substituted or unsubstituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl, or two R.sup.Lb groups are joined to form a substituted or unsubstituted carbocyclic or substituted or unsubstituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form a substituted or unsubstituted heterocyclic ring.
[0225] In certain embodiments, linker L is —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, —NR.sup.LaS(═O).sub.2—, a peptide, a cleavable linker, or an optionally substituted C.sub.1-10 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is replaced with —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—, wherein each instance of R.sup.La is independently hydrogen, optionally substituted C.sub.1-10 alkyl, or a nitrogen protecting group, and wherein each occurrence of R.sup.Lb is independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl, or two R.sup.Lb groups are joined to form an optionally substituted carbocyclic or optionally substituted heterocyclic ring, or R.sup.La and R.sup.Lb are joined to form an optionally substituted heterocyclic ring.
[0226] In certain embodiments, L is an optionally substituted C.sub.1-10 hydrocarbon chain, optionally wherein one or more carbon units of the hydrocarbon chain is replaced with —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—. In certain embodiments, at least one instance of L is a peptide. In certain embodiments, L is a peptide consisting of between 2 and 50, between 2 and 30, between 2 and 20, between 2 and 10, between 2 and 5, between 5 and 50, between 5 and 30, between 5 and 20, between 5 and 10, between 10 and 50, between 10 and 30, between 10 and 20, between 20 and 50, between 20 and 30, or between 30 and 50, inclusive, amino acids (e.g., natural amino acids). In certain embodiments, at least one instance of L is cleavable linker. In certain embodiments, L is a photo-cleavable linker. In certain embodiments, L is not a photo-cleavable linker. In certain embodiments, L is not a photo-cleavable linker. In certain embodiments, L is a linker cleavable by hydrolysis (e.g., under physiological conditions). In certain embodiments, L is a linker cleavable by reduction. In certain embodiments, at least one instance of L is a polymer (e.g., a polymer whose Mw is between 2,000 and 1,000,000, g/mol, inclusive).
[0227] In certain embodiments, at least one instance of L is a substituted or unsubstituted C.sub.1-30 hydrocarbon chain (e.g., substituted or unsubstituted C.sub.6-20 hydrocarbon chain), wherein one or more (e.g., two, three, four, five, or six) carbon units of the hydrocarbon chain is independently replaced with substituted or unsubstituted phenyl, substituted or unsubstituted triazolyl, —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—.
[0228] In certain embodiments, at least one instance of L is a substituted or unsubstituted C.sub.1-30 hydrocarbon chain (e.g., substituted or unsubstituted C.sub.6-20 hydrocarbon chain), wherein:
[0229] one or more (e.g., two) carbon units of the hydrocarbon chain is replaced with substituted or unsubstituted triazolyl (e.g.,
##STR00034##
wherein the nitrogen atom labelled with “*” is attached (e.g., directly or indirectly) to M); and
[0230] optionally one or more (e.g., two, three, four, or five) carbon units of the hydrocarbon chain is independently replaced with substituted or unsubstituted phenyl, —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—.
[0231] In certain embodiments, at least one instance of L is a substituted or unsubstituted C.sub.1-30 hydrocarbon chain (e.g., substituted or unsubstituted C.sub.6-20 hydrocarbon chain), wherein: one or more (e.g., two) carbon units of the hydrocarbon chain is replaced with substituted or unsubstituted triazolyl (e.g.,
##STR00035##
wherein the nitrogen atom labelled with “*” is attached (e.g., directly or indirectly) to M);
[0232] one or more (e.g., two) carbon units of the hydrocarbon chain is replaced with substituted or unsubstituted phenyl; and
[0233] optionally one or more (e.g., two, three, or four) carbon units of the hydrocarbon chain is independently replaced with —O—, —S—, —NR.sup.La—, —NR.sup.LaC(═O)—, —C(═O)NR.sup.La—, —SC(═O)—, —C(═O)S—, —OC(═O)—, —C(═O)O—, —OC(═O)O—, —OC(═O)NR.sup.La—, —NR.sup.LaC(═O)O—, trans-CR.sup.Lb═CR.sup.Lb—, cis-CR.sup.Lb═CR.sup.Lb—, —C≡C—, —OC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2O—, —NR.sup.LaC(R.sup.Lb).sub.2—, —C(R.sup.Lb).sub.2NR.sup.La—, —S(═O).sub.2O—, —OS(═O).sub.2—, —S(═O).sub.2NR.sup.La—, or —NR.sup.LaS(═O).sub.2—.
[0234] In certain embodiments, when at least one instance of L is a substituted C.sub.1-30 hydrocarbon chain (e.g., substituted C.sub.1-10 hydrocarbon chain), optionally wherein one or more carbon units of the hydrocarbon chain is independently replaced as described herein, the substituents on the C.sub.1-30 hydrocarbon chain (e.g., C.sub.1-10 hydrocarbon chain) are independently halogen or substituted or unsubstituted C.sub.1-6 alkyl. In certain embodiments, when at least one instance of L is a substituted C.sub.1-30 hydrocarbon chain (e.g., substituted C.sub.1-10 hydrocarbon chain), optionally wherein one or more carbon units of the hydrocarbon chain is independently replaced as described herein, the substituents on the C.sub.1-30 hydrocarbon chain (e.g., C.sub.1-10 hydrocarbon chain) are independently halogen or unsubstituted C.sub.1-6 alkyl.
[0235] In certain embodiments, L is of one of the following formulae:
##STR00036##
wherein:
[0236] each instance of the carbon atom labeled with “*” is attached to M;
[0237] each instance of R.sup.E is independently halogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, —OR.sup.a, —N(R.sup.a).sub.2, —SR.sup.a, —CN, —SCN, —C(═NR.sup.a)R.sup.a, —C(═NR.sup.a)OR.sup.a, —C(═NR.sup.a)N(R.sup.a).sub.2, —C(═O)R.sup.a, —C(═O)OR.sup.a, —C(═O)N(R.sup.a).sub.2, —N.sub.02, —N(R.sup.a)C(═O)R.sup.a, —N(R.sup.a)C(═O)OR.sup.a, —N(R.sup.a)C(═O)N(R.sup.a).sub.2, —N(R.sup.a)S(═O)R.sup.a, —N(R.sup.a)S(═O)OR.sup.a, —N(R.sup.a)S(═O)N(R.sup.a).sub.2, —N(R.sup.a)S(═O).sub.2R.sup.a, —N(R.sup.a)S(═O).sub.2OR.sup.a, —N(R.sup.a)S(═O).sub.2N(R.sup.a).sub.2, —OC(═O)R.sup.a, —OC(═O)OR.sup.a, or —OC(═O)N(R.sup.a).sub.2;
[0238] each instance of R.sup.a is independently hydrogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, or a sulfur protecting group when attached to a sulfur atom, or two instances of R.sup.a are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring;
[0239] each instance of k is independently 0, 1, 2, 3, or 4;
[0240] each instance of p is independently an integer from 1 to 10, inclusive;
[0241] each instance of q is independently an integer from 1 to 10, inclusive;
[0242] each instance of g is independently an integer from 1 to 10, inclusive; and
[0243] each instance of h is independently an integer from 1 to 10, inclusive.
[0244] In certain embodiments, each instance of L is the same. In certain embodiments, at least two instances of L are different from each other.
[0245] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-a):
##STR00037##
[0246] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-b):
##STR00038##
[0247] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-c):
##STR00039##
[0248] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-d):
##STR00040##
[0249] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-e):
##STR00041##
[0250] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-f):
##STR00042##
[0251] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-g):
##STR00043##
[0252] In certain embodiments, the macromonomer of Formula (III) is of Formula (III-h):
##STR00044##
[0253] As generally defined herein, each instance of M is independently hydrogen or an agent. In certain embodiments, the agent is a small molecule (optionally wherein the molecular weight of the small molecule is not more than 2,000 g/mol). In certain embodiments, the agent is a protein or peptide. In certain embodiments, the agent is a polynucleotide. In certain embodiments, each instance of M is independently hydrogen or a pharmaceutical agent. In certain embodiments, M is hydrogen or a therapeutic, diagnostic, or prophylactic agent. In certain embodiments, M is hydrogen. In certain embodiments, M is not hydrogen. In certain embodiments, at least one instance of M is a pharmaceutical agent. In certain embodiments, M is a therapeutic, diagnostic, or prophylactic agent. In certain embodiments, M is a therapeutic agent. Examples of therapeutic moieties include, but are not limited to, antimicrobial agents, analgesics, antiinflammatory agents, counterirritants, coagulation modifying agents, diuretics, sympathomimetics, anorexics, antacids and other gastrointestinal agents; antiparasitics, antidepressants, antihypertensives, anticholinergics, stimulants, antihormones, central and respiratory stimulants, drug antagonists, lipid-regulating agents, uricosurics, cardiac glycosides, electrolytes, ergot and derivatives thereof, expectorants, hypnotics and sedatives, antidiabetic agents, dopaminergic agents, antiemetics, muscle relaxants, para-sympathomimetics, anticonvulsants, antihistamines, beta-blockers, purgatives, antiarrhythmics, contrast materials, radiopharmaceuticals, antiallergic agents, tranquilizers, vasodilators, antiviral agents, and antineoplastic or cytostatic agents or other agents with anticancer properties, or a combination thereof. Other suitable therapeutic moieties include contraceptives and vitamins as well as micro- and macronutrients. Still other examples include antiinfectives such as antibiotics and antiviral agents; analgesics and analgesic combinations; anorexics; antiheimintics; antiarthritics; antiasthmatic agents; anticonvulsants; antidepressants; antidiuretic agents; antidiarrleals; antihistamines; antiinflammatory agents; antimigraine preparations; antinauseants; antineoplastics; antiparkinsonism drugs; antipruritics; antipsychotics; antipyretics, antispasmodics; anticholinergics; sympathomimetics; xanthine derivatives; cardiovascular preparations including calcium channel blockers and beta-blockers such as pindolol and antiarrhythmics; antihypertensives; diuretics; vasodilators including general coronary, peripheral and cerebral; central nervous system stimulants; cough and cold preparations, including decongestants; hormones such as estradiol and other steroids, including corticosteroids; hypnotics; immunosuppressives; muscle relaxants; parasympatholytics; psychostimulants; sedatives; and tranquilizers; and naturally derived or genetically engineered proteins, polysaccharides, glycoproteins, or lipoproteins. In certain embodiments, at least one instance of M is an anti-cancer agent. In certain embodiments, M is an anti-cancer agent. Anti-cancer agents encompass biotherapeutic anti-cancer agents as well as chemotherapeutic agents. Exemplary biotherapeutic anti-cancer agents include, but are not limited to, interferons, cytokines (e.g., tumor necrosis factor, interferon ct, interferon γ), vaccines, hematopoietic growth factors, monoclonal serotherapy, immunostimulants and/or immunodulatory agents (e.g., IL-1, 2, 4, 6, or 12), immune cell growth factors (e.g., GM-CSF) and antibodies (e.g., HERCEPTIN (trastuzumab), T-DM1, AVASTIN (bevacizumab), ERBITUX (cetuximab), VECTIBIX (panitumumab), RITUXAN (rituximab), BEXXAR (tositumomab)). Exemplary chemotherapeutic agents include, but are not limited to, anti-estrogens (e.g., tamoxifen, raloxifene, and megestrol), LHRH agonists (e.g., goscrclin and leuprolide), anti-androgens (e.g., flutamide and bicalutamide), photodynamic therapies (e.g., vertoporfin (BPD-MA), phthalocyanine, photosensitizer Pc4, and demethoxy-hypocrellin A (2BA-2-DMHA)), nitrogen mustards (e.g., cyclophosphamide, ifosfamide, trofosfamide, chlorambucil, estramustine, and melphalan), nitrosoureas (e.g., carmustine (BCNU) and lomustine (CCNU)), alkylsulphonates (e.g., busulfan and treosulfan), triazenes (e.g., dacarbazine, temozolomide), platinum containing compounds (e.g., cisplatin, carboplatin, oxaliplatin), vinca alkaloids (e.g., vincristine, vinblastine, vindesine, and vinorelbine), taxoids (e.g., paclitaxel or a paclitaxel equivalent) docosahexaenoic acid bound-paclitaxel (DHA-paclitaxel, Taxoprexin), polyglutamate bound-paclitaxel (PG-paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX), the tumor-activated prodrug (TAP) ANG1005 (Angiopep-2 bound to three molecules of paclitaxel), paclitaxel-EC-1 (paclitaxel bound to the erbB2-recognizing peptide EC-1), and glucose-conjugated paclitaxel, e.g., 2′-paclitaxel methyl 2-glucopyranosyl succinate; docetaxel, taxol), epipodophyllins (e.g., etoposide, etoposide phosphate, teniposide, topotecan, 9-aminocamptothecin, camptoirinotecan, irinotecan, crisnatol, mytomycin C), anti-metabolites, DHFR inhibitors (e.g., methotrexate, dichloromethotrexate, trimetrexate, edatrexate), IMP dehydrogenase inhibitors (e.g., mycophenolic acid, tiazofurin, ribavirin, and EICAR), ribonuclotide reductase inhibitors (e.g. hydroxyurea and deferoxamine), uracil analogs (e.g., 5-fluorouracil (5-FU), floxuridine, doxifluridine, ratitrexed, tegafur-uracil, capecitabine), cytosine analogs (e.g., cytarabine (ara C), cytosine arabinoside, and fludarabine), purine analogs (e.g., mercaptopurine and Thioguanine), Vitamin D3 analogs (e.g., EB 1089, CB 1093, and KH 1060), isoprenylation inhibitors (e.g., lovastatin), dopaminergic neurotoxins (e.g., 1-methyl-4-phenylpyridinium ion), cell cycle inhibitors (e.g., staurosporine), actinomycin (e.g., actinomycin D, dactinomycin), bleomycin (e.g., bleomycin A2, bleomycin B2, peplomycin), anthracycline (e.g., daunorubicin, doxorubicin, pegylated liposomal doxorubicin, idarubicin, epirubicin, pirarubicin, zorubicin, mitoxantrone), MDR inhibitors (e.g., verapamil), Ca.sup.2+ ATPase inhibitors (e.g., thapsigargin), imatinib, thalidomide, lenalidomide, tyrosine kinase inhibitors (e.g., axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTIN™, AZD2171), dasatinib (SPRYCEL®, BMS-354825), erlotinib (TARCEVA®), gefitinib (IRESSA®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib (TYKERB®, TYVERB®), lestaurtinib (CEP-701), neratinib (HKI-272), nilotinib (TASIGNA®), semaxanib (semaxinib, SU5416), sunitinib (SUTENT®, SU11248), toceranib (PALLADIA®), vandetanib (ZACTIMA®, ZD6474), vatalanib (PTK787, PTK/ZK), trastuzumab (HERCEPTIN®), bevacizumab (AVASTIN®), rituximab (RITUXAN®), cetuximab (ERBITUX®), panitumumab (VECTIBIX®), ranibizumab (Lucentis®), nilotinib (TASIGNA®), sorafenib (NEXAVAR®), everolimus (AFINITOR®), alemtuzumab (CAMPATH®), gemtuzumab ozogamicin (MYLOTARG®), temsirolimus (TORISEL®), ENMD-2076, PCI-32765, AC220, dovitinib lactate (TKI258, CHIR-258), BIBW 2992 (TOVOK™), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC1-2036, BMS-690154, CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, and/or XL228), proteasome inhibitors (e.g., bortezomib (VELCADE)), mTOR inhibitors (e.g., rapamycin, temsirolimus (CCI-779), everolimus (RAD-001), ridaforolimus, AP23573 (Ariad), AZD8055 (AstraZeneca), BEZ235 (Novartis), BGT226 (Norvartis), XL765 (Sanofi Aventis), PF-4691502 (Pfizer), GDC0980 (Genetech), SF1126 (Semafoe), and OSI-027 (OSI)), oblimersen, gemcitabine, carminomycin, leucovorin, pemetrexed, cyclophosphamide, dacarbazine, procarbizine, prednisolone, dexamethasone, campathecin, plicamycin, asparaginase, aminopterin, methopterin, porfiromycin, melphalan, leurosidine, leurosine, chlorambucil, trabectedin, procarbazine, discodermolide, carminomycin, aminopterin, and hexamethyl melamine. In certain embodiments, at least one instance of M is camptothecin. In certain embodiments, at least one instance of M is IRT. In certain embodiments, at least one instance of M is SN-38. In certain embodiments, at least one instance of M is 5-FU. In certain embodiments, at least two instances of M are camptothecin and 5-FU. In certain embodiments, at least two instances of M are IRT and 5-FU. In certain embodiments, at least two instances M are SN-38 and 5-FU. In certain embodiments, at least three instances of M are IRT, 5-FU, and oxaliplatin or a derivative thereof. In certain embodiments, at least three instances of M are CPT, 5-FU, and oxaliplatin or a derivative thereof. In certain embodiments, at least three instances of M are SN-38, 5-FU, and oxaliplatin or a derivative thereof.
[0254] In certain embodiments, M is a diagnostic agent. Exemplary diagnostic agents include, but are not limited to, fluorescent molecules; gases; metals; commercially available imaging agents used in positron emissions tomography (PET), computer assisted tomography (CAT), single photon emission computerized tomography, x-ray, fluoroscopy, and magnetic resonance imaging (MRI); and contrast agents. Examples of suitable materials for use as contrast agents in MRI include gadolinium chelates, as well as iron, magnesium, manganese, copper, and chromium. Examples of materials useful for CAT and x-ray imaging include iodine-based materials. In certain embodiments, the diagnostic agent is used in magnetic resonance imaging (MRI), such as iron oxide particles or gadolinium complexes. Gadolinium complexes that have been approved for clinical use include gadolinium chelates with DTPA, DTPA-BMA, DOTA and HP-DO3A which are reviewed in Aime, et al. (Chemical Society Reviews (1998), 27:19-29), the entire teachings of which are incorporated herein by reference.
[0255] In certain embodiments, M is a prophylactic agent. Prophylactic agents that can be included in the conjugates of the invention include, but are not limited to, antibiotics, nutritional supplements, and vaccines. Vaccines may comprise isolated proteins or peptides, inactivated organisms and viruses, dead organisms and viruses, genetically altered organisms or viruses, and cell extracts. Prophylactic agents may be combined with interleukins, interferon, cytokines, and adjuvants such as cholera toxin, alum, Freund's adjuvant.
[0256] In certain embodiments, each instance of M is the same. In certain embodiments, at least two instances of M are different from each other. In certain embodiments, at least three instances of M are different from each other. In certain embodiments, at least four instances of M are different from each other.
[0257] M can be conjugated to the macromonomer using any suitable conjugation technique. For instance, EDC-NHS chemistry (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide), or a reaction involving a maleimide or a carboxylic acid, which can be conjugated to one end of a thiol, an amine, or a similarly functionalized polyether. The conjugation can be performed in an organic solvent, such as, but not limited to, methylene chloride, acetonitrile, chloroform, dimethylformamide, tetrahydrofuran, acetone, or the like. Specific reaction conditions can be determined by those of ordinary skill in the art using no more than routine experimentation.
[0258] In another set of embodiments, a conjugation reaction may be performed by reacting the agent that includes a hydroxyl, thiol, or amino group with a polymer comprising a carboxylic acid functional group. Such a reaction may occur as a single-step reaction, i.e., the conjugation is performed with or without using intermediates such as N-hydroxysuccinimide or a maleimide. The conjugation reaction between the amine-containing, thiol-containing, or hydroxyl-containing moiety and the carboxylic acid-terminated polymer may be achieved in one embodiment, by adding the amine-containing, thiol-containing, or hydroxyl-containing moiety, solubilized in an organic solvent such as, but not limited to, dichloromethane, acetonitrile, chloroform, tetrahydrofuran, acetone, formamide, dimethylformamide, pyridines, dioxane, or dimethysulfoxide, to a solution containing the carboxylic acid-terminated polymer. The carboxylic acid-terminated polymer may be contained within an organic solvent such as, but not limited to, dichloromethane, acetonitrile, chloroform, dimethylformamide, tetrahydrofuran, or acetone. Reaction between the amine-containing moiety and the carboxylic acid-terminated polymer may occur spontaneously in some cases. Unconjugated macromonomers may be washed away after such reactions, and the polymer may be precipitated in solvents such as, for instance, ethyl ether, hexane, methanol, or ethanol.
[0259] The synthesis of Pt-BASPs involves ROMP of MM in step (a) and ROMP of the platinum complex in step (b). In certain embodiments, the ROMP catalyst is a tungsten (W), molybdenum (Mo), or ruthenium (Ru) catalyst. In certain embodiments, the ROMP catalyst is a ruthenuim catalyst. ROMP catalysts useful in the synthetic methods described herein include catalysts as depicted below, and as described in Grubbs et al., Acc. Chem. Res. 1995, 28, 446-452; U.S. Pat. No. 5,811,515; Schrock et al., Organometallics (1982) 1 1645; Gallivan et al., Tetrahedron Letters (2005) 46:2577-2580; Furstner et al., J. Am. Chem. Soc. (1999) 121:9453; and Chem. Eur. J. (2001) 7:5299; the entire contents of each of which are incorporated herein by reference.
[0260] In certain embodiments, the ROMP catalyst is a Grubbs catalyst. In certain embodiments, the Grubbs catalyst is selected from the group consisting of:
##STR00045## [0261] Benzylidenebis-(tricyclohexylphosphine)-dichlororuthenium (X=Cl); [0262] Benzylidenebis-(tricyclohexylphosphine)-dibromoruthenium (X=Br); [0263] Benzylidenebis-(tricyclohexylphosphine)-diiodoruthenium (X=I);
##STR00046## [0264] 1,3-(Bis(mesityl)-2-imidazolidinylidene)dichloro-(phenylmethylene) (tricyclohexyl-phosphine)ruthenium (X=Cl; R=cyclohexyl); [0265] 1,3-(Bis(mesityl)-2-imidazolidinylidene)dibromo-(phenylmethylene) (tricyclohexyl-phosphine)ruthenium (X=Br; R=cyclohexyl); [0266] 1,3-(Bis(mesityl)-2-imidazolidinylidne) o-penylmethylene) (tricyclohexyl-phosphine)ruthenium (X=I; R=cyclohexyl); [0267] 1,3-(Bis(mesityl)-2-imidazolidinylidene)dichloro-(phenylmethylene) (triphenylphosphine)ruthenium (X=Cl; R=phenyl); [0268] 1,3-(Bis(mesityl)-2-imidazolidinylidene)dichloro-(phenylmethylene) (tribenzylphosphine)ruthenium (X=Cl; R=benzyl);
##STR00047##
[0269] In certain embodiments, the ROMP catalyst is a Grubbs-Hoveyda catalyst. In certain embodiments, the Grubbs-Hoveyda catalyst is selected from the group consisting of:
##STR00048##
[0270] In certain embodiments, the ROMP catalyst is selected from the group consisting of:
##STR00049##
[0271] In certain embodiments, the ROMP catalyst is of the formula:
##STR00050##
[0272] The ROMP can be conducted in one or more aprotic solvents. The term “aprotic solvent” means a non-nucleophilic solvent having a boiling point range above ambient temperature, preferably from about 25° C. to about 190° C. at atmospheric pressure. In certain embodiments, the aprotic solvent has a boiling point from about 80° C. to about 160° C. at atmospheric pressure. In certain embodiments, the aprotic solvent has a boiling point from about 80° C. to about 150° C. at atmospheric pressure. Examples of such solvents are methylene chloride, acetonitrile, toluene, DMF, diglyme, THF, and DMSO.
[0273] The ROMP can be quenched with a vinyl ether of the formula
##STR00051##
Each of R.sup.V1, R.sup.V2, R.sup.V3, and R.sup.V4 is independently optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted carbocyclyl, optionally substituted phenyl, optionally substituted heterocyclyl, or optionally substituted heteroaryl. In certain embodiments, R.sup.V1 is optionally substituted alkyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is unsubstituted alkyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is substituted alkyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is methyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is ethyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is propyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is optionally substituted alkenyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is unsubstituted alkenyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, R.sup.V1 is vinyl, and R.sup.V2, R.sup.V3, and R.sup.V4 are hydrogen. In certain embodiments, at least one of R.sup.V1, R.sup.V2, R.sup.V3, and R.sup.V4 is conjugated with a diagnostic agent as defined above. In certain embodiments, the ROMP is quenched by ethyl vinyl ether. Excess ethyl vinyl ether can be removed from the Pt-BASPs by vacuum.
[0274] The Pt-BASPs can be multi-agent loaded star polymers. In certain embodiments, the inventive Pt-BASPs are capable of releasing multiple chemotherapeutic agents for combination therapy. In certain embodiments, the Pt-BASPs are capable of releasing two chemotherapeutic agents. In certain embodiments, the Pt-BASPs are capable of releasing three chemotherapeutic agents. In certain embodiments, the Pt-BASPs are capable of releasing four chemotherapeutic agents. In certain embodiments, the Pt-BASPs are capable of releasing five chemotherapeutic agents. In certain embodiments, the Pt-BASPs incorporate only a platinum-based agent which is introduced from the platinum-based complex crosslinkers. In certain embodiments, the inventive Pt-BASPs incorporate only cisplatin. In certain embodiments, the Pt-BASPs incorporate one or more therapeutic, diagnostic, or prophylactic agents. The one or more therapeutic, diagnostic, or prophylactic agents are introduced from macromonomers in the synthesis of Pt-BASPs. In certain embodiments, a therapeutic agent is incorporated in the macromonomer. In certain embodiments, an anti-cancer agent is incorporated in the macromonomer. In certain embodiments, the macromonomer is camptothecin (CPT) macromonomer. In certain embodiments, the macromonomer is IRT macromonomer. In certain embodiments, the macromonomer is 5-FU macromonomer. In certain embodiments, the macromonomer is SN-38 macromonomer. In certain embodiments, the macromonomer is doxorubicin (DOX) macromonomer. In certain embodiments, the inventive Pt-BASPs incorporate oxaliplatin or a derivative thereof and CPT. In certain embodiments, the inventive Pt-BASPs incorporate oxaliplatin or a derivative thereof and IRT. In certain embodiments, the inventive Pt-BASPs incorporate oxaliplatin or a derivative thereof and 5-FU. In certain embodiments, the inventive Pt-BASPs incorporate an oxaliplatin prodrug, CPT, and 5-FU. In certain embodiments, the inventive Pt-BASPs incorporate an oxaliplatin prodrug, IRT, and 5-FU. In certain embodiments, the inventive Pt-BASPs incorporate an oxaliplatin prodrug, SN-38, and 5-FU. In certain embodiments, the one or more therapeutic, diagnostic, or prophylactic agents are connected to Pt-BASPs by a photocleavable linker. In certain embodiments, the inventive Pt-BASPs incorporate a diagnostic agent and a platinum therapeutic agent. In certain embodiments, the inventive Pt-BASPs incorporate a prophylactic agent and a platinum therapeutic agent.
[0275] In some cases, the Pt-BASPs are of the form of nanoparticles, i.e., the particle have a characteristic dimension of less than about 1 micrometer, where the characteristic dimension of a particle is the diameter of a perfect sphere having the same volume as the particle. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 300 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 200 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 150 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 100 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 50 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 30 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 20 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension of less than about 10 nm. In certain embodiments, the Pt-BASP particle has a characteristic dimension between 6 and 250 nm, inclusive. In certain embodiments, the Pt-BASP particle has a characteristic dimension between 8 and 200 nm, inclusive. In certain embodiments, the Pt-BASP particle has a characteristic dimension between 12 and 200 nm, inclusive. In certain embodiments, the Pt-BASP particle has a characteristic dimension between 50 and 200 nm, inclusive.
[0276] In certain embodiments, the Pt-BASPs are biodegradable, i.e., the polymer is able to degrade, chemically and/or biologically, within a physiological environment, such as within the body. For instance, the polymer may be one that hydrolyzes spontaneously upon exposure to water (e.g., within a subject), the polymer may degrade upon exposure to heat (e.g., at temperatures of about 37° C.). Degradation of a polymer may occur at varying rates, depending on the polymer or copolymer used. For example, the half-life of the polymer (the time at which 50% of the polymer is degraded into monomers and/or other nonpolymeric moieties) may be on the order of days, weeks, months, or years, depending on the polymer. The polymers may be biologically degraded, e.g., by enzymatic activity or cellular machinery, in some cases, for example, through exposure to a lysozyme (e.g., having relatively low pH). In some cases, the polymers may be broken down into monomers and/or other nonpolymeric moieties that cells can either reuse or dispose of without significant toxic effect on the cells (for example, polylactide may be hydrolyzed to form lactic acid, polyglycolide may be hydrolyzed to form glycolic acid, etc.).
[0277] In certain embodiments, the solvents in step (a) is the same as the solvent of the platinum complex solution in step (b). In certain embodiments, the solvents in step (a) is different from the solvent of the platinum complex solution in step (b). Exemplary solvents for step (a) and platinum complex solution include, but are not limited to, methylene chloride, acetonitrile, chloroform, dimethylformamide, tetrahydrofuran, and acetone. Specific reaction conditions can be determined by those of ordinary skill in the art using no more than routine experimentation.
[0278] In certain embodiments of one type of macromonomer in Pt-BASPs, the amount of all the macromonomers, the platinum complex crosslinker, and the metathesis catalyst is of the molar ratio m:N:1, wherein m is an integer from 1 to 20, inclusive; and N is an integer from 1 to 20, inclusive. In certain embodiments, m is an integer from 3 to 12 inclusive. In certain embodiments, m is 1. In certain embodiments, m is 2. In certain embodiments, m is 3. In certain embodiments, m is 4. In certain embodiments, m is 5. In certain embodiments, m is 6. In certain embodiments, m is 7. In certain embodiments, m is 8. In certain embodiments, m is 9. In certain embodiments, m is 10. In certain embodiments, m is 11. In certain embodiments, m is 12. In certain embodiments, N is an integer from 1 to 10 inclusive. In certain embodiments, N is 1. In certain embodiments, N is 2. In certain embodiments, N is 3. In certain embodiments, N is 4. In certain embodiments, N is 5. In certain embodiments, N is 6. In certain embodiments, N is 7. In certain embodiments, N is 8. In certain embodiments, N is 9. In certain embodiments, N is 10. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the ratio 5:3:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 5:5:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 5:7:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 7:3:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 7:5:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 7:7:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 11:1:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 11:3:1. In certain embodiments, the amount of macromonomer, platinum complex crosslinker, and metathesis catalyst is of the molar ratio 11:5:1.
[0279] In certain embodiments, the molar ratio of the first macromonomer to the metathesis catalyst is between 1:1 to 20:1, inclusive; the molar ratio of the second macromonomer to the metathesis catalyst is between 1:1 to 20:1, inclusive; and the molar ratio of the platinum complex to the metathesis catalyst is between 1:1 to 20:1, inclusive. In certain embodiments, the molar ratio of the first macromonomer to the metathesis catalyst is between 1:1 to 10:1, inclusive; the molar ratio of the second macromonomer to the metathesis catalyst is between 1:1 to 10:1, inclusive; and the molar ratio of the platinum complex to the metathesis catalyst is between 1:1 to 10:1, inclusive. In certain embodiments, the molar ratio of the first macromonomer to the metathesis catalyst is between 1:1 to 5:1, inclusive; the molar ratio of the second macromonomer to the metathesis catalyst is between 1:1 to 5:1, inclusive; and the molar ratio of the platinum complex to the metathesis catalyst is between 1:1 to 5:1, inclusive.
[0280] In certain embodiments of multi-agent loaded Pt-BASPs, the amount of the first macromonomer, second macromonomer, and platinum complex crosslinker is of the molar ratio of m1:m2:N, wherein m1 and m2 are each independently an integer from 1 to 20, inclusive; and N is an integer from 1 to 20, inclusive. In certain embodiments, the amount of the first macromonomer, second macromonomer, and platinum complex crosslinker is of the molar ratio of 3:4:3. In certain embodiments, the amount of the first macromonomer, second macromonomer, and platinum complex crosslinker is of the molar ratio of 4:3:3. In certain embodiments of more than one types of macromonomers in the multi-agent loaded Pt-BASPs, the amount of the first macromonomer, second macromonomer, and platinum complex crosslinker is of the molar ratio of m1:m2:N: 1, wherein m1 and m2 are each independently an integer from 1 to 20, inclusive; and N is an integer from 1 to 20, inclusive. In certain embodiments, the amount of the first macromonomer, second macromonomer, and platinum complex crosslinker is of the molar ratio of 3:4:3:1. In certain embodiments, the amount of the first macromonomer, second macromonomer, and platinum complex crosslinker is of the molar ratio of 4:3:3:1.
[0281] Exposure to physiologically relevant conditions can lead to the Pt-BASPs degradation and controlled, extended release of platinum-based agents. In certain embodiments, the release rate can increased by addition of GSH. In vitro cytotoxicity assays demonstrates that Pt-BASPs effectively kill cancer cells (
Pharmaceutical Compositions, Kits, and Administration
[0282] The present invention provides pharmaceutical compositions comprising Pt-BASPs, as described herein, and optionally a pharmaceutically acceptable excipient. In certain embodiments, the Pt-BASPs are provided in an effective amount in the pharmaceutical composition. In certain embodiments, the effective amount is a therapeutically effective amount. In certain embodiments, the effective amount is a prophylactically effective amount.
[0283] Pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. In general, such preparatory methods include the steps of bringing the Pt-BASPs into association with a carrier and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping and/or packaging the product into a desired single- or multi-dose unit.
[0284] Pharmaceutical compositions can be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses. A “unit dose” is a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject and/or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.
[0285] Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w/w) active ingredient.
[0286] Pharmaceutically acceptable excipients used in the manufacture of provided pharmaceutical compositions include inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents may also be present in the composition.
[0287] Exemplary diluents include calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.
[0288] Exemplary granulating and/or dispersing agents include potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation-exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethyl cellulose, magnesium aluminum silicate (VEEGUM), sodium lauryl sulfate, quaternary ammonium compounds, and mixtures thereof.
[0289] Exemplary surface active agents and/or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxy polymethylene, polyacrylic acid, acrylic acid polymer, and carboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g., carboxymethylcellulose sodium, powdered cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween® 20), polyoxyethylene sorbitan (Tween® 60), polyoxyethylene sorbitan monooleate (Tween® 80), sorbitan monopalmitate (Span® 40), sorbitan monostearate (Span® 60), sorbitan tristearate (Span® 65), glyceryl monooleate, sorbitan monooleate (Span® 80)), polyoxyethylene esters (e.g., polyoxyethylene monostearate (MYRJ 45), polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor™), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (BRIJ 30)), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, PLURONIC F-68, Poloxamer-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and/or mixtures thereof.
[0290] Exemplary binding agents include starch (e.g., cornstarch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husks, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate (VEEGUM), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.
[0291] Exemplary preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.
[0292] Exemplary antioxidants include alpha tocopherol, ascorbic acid, acorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite.
[0293] Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodium edetate, trisodium edetate, calcium disodium edetate, dipotassium edetate, and the like), citric acid and salts and hydrates thereof (e.g., citric acid monohydrate), fumaric acid and salts and hydrates thereof, malic acid and salts and hydrates thereof, phosphoric acid and salts and hydrates thereof, and tartaric acid and salts and hydrates thereof. Exemplary antimicrobial preservatives include benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal.
[0294] Exemplary antifungal preservatives include butyl paraben, methyl paraben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid.
[0295] Exemplary alcohol preservatives include ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol.
[0296] Exemplary acidic preservatives include vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.
[0297] Other preservatives include tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisol (BHA), butylated hydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, GLYDANT PLUS, PHENONIP, methylparaben, GERMALL 115, GERMABEN II, NEOLONE, KATHON, and EUXYL.
[0298] Exemplary buffering agents include citrate buffer solutions, acetate buffer solutions, phosphate buffer solutions, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and mixtures thereof.
[0299] Exemplary lubricating agents include magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and mixtures thereof.
[0300] Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, camomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed, geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, sasquana, savoury, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germ oils. Exemplary synthetic oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and mixtures thereof.
[0301] Liquid dosage forms for oral and parenteral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. In certain embodiments for parenteral administration, the conjugates of the invention are mixed with solubilizing agents such as Cremophor™, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and mixtures thereof.
[0302] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions can be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation can be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.
[0303] The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
[0304] Compositions for rectal or vaginal administration are typically suppositories which can be prepared by mixing the conjugates of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active ingredient.
[0305] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium compounds, (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may include a buffering agent.
[0306] Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally comprise opacifying agents and can be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. Solid compositions of a similar type can be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polethylene glycols and the like.
[0307] Dosage forms for topical and/or transdermal administration of a compound of this invention may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and/or patches. Generally, the Pt-BASPs are admixed under sterile conditions with a pharmaceutically acceptable carrier and/or any needed preservatives and/or buffers as can be required. Additionally, the present invention contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of an active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and/or dispensing the active ingredient in the proper medium. Alternatively or additionally, the rate can be controlled by either providing a rate controlling membrane and/or by dispersing the active ingredient in a polymer matrix and/or gel.
[0308] Suitable devices for use in delivering intradermal pharmaceutical compositions described herein include short needle devices such as those described in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositions can be administered by devices which limit the effective penetration length of a needle into the skin, such as those described in PCT publication WO 99/34850 and functional equivalents thereof. Jet injection devices which deliver liquid vaccines to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable. Jet injection devices are described, for example, in U.S. Pat. Nos. 5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189; 5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335; 5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880; 4,940,460; and PCT publications WO 97/37705 and WO 97/13537. Ballistic powder/particle delivery devices which use compressed gas to accelerate the compound in powder form through the outer layers of the skin to the dermis are suitable. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration.
[0309] Formulations suitable for topical administration include, but are not limited to, liquid and/or semi liquid preparations such as liniments, lotions, oil-in-water and/or water-in-oil emulsions such as creams, ointments, and/or pastes, and/or solutions and/or suspensions. Topically-administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient can be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further comprise one or more of the additional ingredients described herein.
[0310] A pharmaceutical composition of the invention can be prepared, packaged, and/or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles of Pt-BASPs described herein. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant can be directed to disperse the powder and/or using a self-propelling solvent/powder dispensing container.
[0311] Low boiling propellants generally include liquid propellants having a boiling point of below 65° F. at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w/w) of the composition, and the active ingredient may constitute 0.1 to 20% (w/w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic and/or solid anionic surfactant and/or a solid diluent (which may have a particle size of the same order as particles comprising the active ingredient).
[0312] Pharmaceutical compositions of the invention formulated for pulmonary delivery may provide the Pt-BASPs in the form of droplets of a solution and/or suspension. Such formulations can be prepared, packaged, and/or sold as aqueous and/or dilute alcoholic solutions and/or suspensions, optionally sterile, comprising the active ingredient, and may conveniently be administered using any nebulization and/or atomization device. Such formulations may further comprise one or more additional ingredients including, but not limited to, a flavoring agent such as saccharin sodium, a volatile oil, a buffering agent, a surface active agent, and/or a preservative such as methylhydroxybenzoate. The droplets provided by this route of administration may have an average diameter in the range from about 0.1 to about 200 nanometers.
[0313] Formulations described herein as being useful for pulmonary delivery are useful for intranasal delivery of a pharmaceutical composition of the invention. Another formulation suitable for intranasal administration is a coarse powder comprising the Pt-BASPs and having an average particle from about 0.2 to 500 micrometers. Such a formulation is administered by rapid inhalation through the nasal passage from a container of the powder held close to the nares.
[0314] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with ordinary experimentation.
[0315] Pt-BASPs provided herein are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of the compositions of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend upon a variety of factors including the disease being treated and the severity of the disorder; the activity of the specific active ingredient employed; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific active ingredient employed; the duration of the treatment; drugs used in combination or coincidental with the specific active ingredient employed; and like factors well known in the medical arts.
[0316] The Pt-BASPs and compositions provided herein can be administered by any route, including enteral (e.g., oral), parenteral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, subcutaneous, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), mucosal, nasal, bucal, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol. Specifically contemplated routes are oral administration, intravenous administration (e.g., systemic intravenous injection), regional administration via blood and/or lymph supply, and/or direct administration to an affected site. In general the most appropriate route of administration will depend upon a variety of factors including the nature of the agent (e.g., its stability in the environment of the gastrointestinal tract), and/or the condition of the subject (e.g., whether the subject is able to tolerate oral administration).
[0317] The exact amount of a compound required to achieve an effective amount will vary from subject to subject, depending, for example, on species, age, and general condition of a subject, severity of the side effects or disorder, identity of the particular compound(s), mode of administration, and the like. The desired dosage can be delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dosage can be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations).
[0318] In certain embodiments, an effective amount of the inventive polymer for administration one or more times a day to a 70 kg adult human may comprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000 mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg, about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1 mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about 1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosage form.
[0319] In certain embodiments, the inventive polymer may be at dosage levels sufficient to deliver from about 0.001 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 50 mg/kg, preferably from about 0.1 mg/kg to about 40 mg/kg, preferably from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, and more preferably from about 1 mg/kg to about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired therapeutic effect.
[0320] It will be appreciated that dose ranges as described herein provide guidance for the administration of provided pharmaceutical compositions to an adult. The amount to be administered to, for example, a child or an adolescent can be determined by a medical practitioner or person skilled in the art and can be lower or the same as that administered to an adult.
[0321] It will be also appreciated that a compound or composition, as described herein, can be administered in combination with one or more additional pharmaceutical agents. The compounds or compositions can be administered in combination with additional pharmaceutical agents that improve their bioavailability, reduce and/or modify their metabolism, inhibit their excretion, and/or modify their distribution within the body. It will also be appreciated that the therapy employed may achieve a desired effect for the same disorder, and/or it may achieve different effects.
[0322] The compound or composition can be administered concurrently with, prior to, or subsequent to, one or more additional pharmaceutical agents, which may be useful as, e.g., combination therapies. Pharmaceutical agents include therapeutically active agents. Pharmaceutical agents also include prophylactically active agents. Each additional pharmaceutical agent may be administered at a dose and/or on a time schedule determined for that pharmaceutical agent. The additional pharmaceutical agents may also be administered together with each other and/or with the compound or composition described herein in a single dose or administered separately in different doses. The particular combination to employ in a regimen will take into account compatibility of the inventive compound with the additional pharmaceutical agents and/or the desired therapeutic and/or prophylactic effect to be achieved. In general, it is expected that the additional pharmaceutical agents utilized in combination be utilized at levels that do not exceed the levels at which they are utilized individually. In some embodiments, the levels utilized in combination will be lower than those utilized individually.
[0323] Exemplary additional pharmaceutical agents include, but are not limited to, anti-proliferative agents, anti-cancer agents, anti-diabetic agents, anti-inflammatory agents, immunosuppressant agents, and a pain-relieving agent. Pharmaceutical agents include small organic molecules such as drug compounds (e.g., compounds approved by the U.S. Food and Drug Administration as provided in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, small molecules linked to proteins, glycoproteins, steroids, nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells.
[0324] Also encompassed by the invention are kits (e.g., pharmaceutical packs). The inventive kits may be useful for treating a proliferative disease (e.g., cancer (e.g., leukemia, melanoma, multiple myeloma), benign neoplasm, angiogenesis, inflammatory disease, autoinflammatory disease, or autoimmune disease). The kits provided may comprise the Pt-BASPs described herein, or a pharmaceutical composition thereof, and a container (e.g., a vial, ampule, bottle, syringe, and/or dispenser package, or other suitable container). In some embodiments, provided kits may optionally further include a second container comprising a pharmaceutical excipient for dilution or suspension of an inventive pharmaceutical composition or compound. In some embodiments, the inventive pharmaceutical composition or compound provided in the container and the second container are combined to form one unit dosage form.
Methods of Treatment and Uses
[0325] The present invention also provides methods of using the Pt-BASPs described herein, or a pharmaceutical composition thereof, for the treatment or prevention of a proliferative disease such as cancer (e.g., lung cancer, large bowel cancer, pancreas cancer, biliary tract cancer, colorectal cancer, or endometrial cancer), benign neoplasm, angiogenesis, inflammatory disease, autoinflammatory disease, or autoimmune disease in a subject.
[0326] In some embodiments, the Pt-BASPs described herein, or a pharmaceutical composition thereof are useful in treating a cancer. In some embodiments, the Pt-BASPs described herein, or a pharmaceutical composition thereof, are useful to delay the onset of, slow the progression of, or ameliorate the symptoms of cancer. In some embodiments, the Pt-BASPs described herein, or a pharmaceutical composition thereof, are administered in combination with other compounds, drugs, or therapeutics to treat cancer.
[0327] In some embodiments, the Pt-BASPs described herein, or a pharmaceutical composition thereof are useful for treating a cancer including, but not limited to, acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangio sarcoma), appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g., cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary cancer, medullary carcinoma of the breast), brain cancer (e.g., meningioma; glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchus cancer, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarcinoma), Ewing sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma), familiar hypereosinophilia, gall bladder cancer, gastric cancer (e.g., stomach adenocarcinoma), gastrointestinal stromal tumor (GIST), head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma (OSCC), throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)), hematopoietic cancers (e.g., leukemia such as acute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicular lymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., “Waldenstrim's macroglobulinemia”), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphomalleukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sezary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); a mixture of one or more leukemia/lymphoma as described above; and multiple myeloma), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease), hemangioblastoma, inflammatory myofibroblastic tumors, immunocytic amyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular cancer (HCC), malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV), essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM), a.k.a. myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g., gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumors), penile cancer (e.g., Paget's disease of the penis and scrotum), pinealoma, primitive neuroectodermal tumor (PNT), prostate cancer (e.g., prostate adenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g., appendix cancer), soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland carcinoma, sweat gland carcinoma, synovioma, testicular cancer (e.g., seminoma, testicular embryonal carcinoma), thyroid cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer), urethral cancer, vaginal cancer and vulvar cancer (e.g., Paget's disease of the vulva).
[0328] In some embodiments, the Pt-BASPs described herein, or a pharmaceutical composition thereof, are useful in treating lung cancer, head-and-neck cancer, esophagus cancer, stomach cancer, breast cancer, pancreas cancer, colorectal cancer, liver cancer, kidney cancer, prostate caner, glioblastomas, metastatic melanomas, peritoneal or pleural mesotheliomas.
[0329] In some embodiments, the proliferative disease is a benign neoplasm. All types of benign neoplasms disclosed herein or known in the art are contemplated as being within the scope of the invention. In some embodiments, the proliferative disease is associated with angiogenesis. All types of angiogenesis disclosed herein or known in the art are contemplated as being within the scope of the invention. In certain embodiments, the proliferative disease is an inflammatory disease. All types of inflammatory diseases disclosed herein or known in the art are contemplated as being within the scope of the invention. In certain embodiments, the inflammatory disease is rheumatoid arthritis. In some embodiments, the proliferative disease is an autoinflammatory disease. All types of autoinflammatory diseases disclosed herein or known in the art are contemplated as being within the scope of the invention. In some embodiments, the proliferative disease is an autoimmune disease. All types of autoimmune diseases disclosed herein or known in the art are contemplated as being within the scope of the invention.
[0330] In certain embodiments, the methods described herein include administering to a subject with an effective amount of the Pt-BASPs described herein, or a pharmaceutical composition thereof. In certain embodiments, the methods described herein include implanting to a subject with an effective amount of the Pt-BASPs described herein, or a pharmaceutical composition thereof.
[0331] In certain embodiments, the Pt-BASPs described herein, or a pharmaceutical composition thereof, are administered in combination with one or more additional pharmaceutical agents described herein. In certain embodiments, the additional pharmaceutical agent is an anti-cancer agent. Anti-cancer agents encompass biotherapeutic anti-cancer agents as well as chemotherapeutic agents. Exemplary biotherapeutic anti-cancer agents include, but are not limited to, interferons, cytokines (e.g., tumor necrosis factor, interferon ct, interferon γ), vaccines, hematopoietic growth factors, monoclonal serotherapy, immunostimulants and/or immunodulatory agents (e.g., IL-1, 2, 4, 6, or 12), immune cell growth factors (e.g., GM-CSF) and antibodies (e.g., HERCEPTIN (trastuzumab), T-DM1, AVASTIN (bevacizumab), ERBITUX (cetuximab), VECTIBIX (panitumumab), RITUXAN (rituximab), BEXXAR (tositumomab)). Exemplary chemotherapeutic agents include, but are not limited to, anti-estrogens (e.g., tamoxifen, raloxifene, and megestrol), LHRH agonists (e.g., goscrclin and leuprolide), anti-androgens (e.g., flutamide and bicalutamide), photodynamic therapies (e.g., vertoporfin (BPD-MA), phthalocyanine, photosensitizer Pc4, and demethoxy-hypocrellin A (2BA-2-DMHA)), nitrogen mustards (e.g., cyclophosphamide, ifosfamide, trofosfamide, chlorambucil, estramustine, and melphalan), nitrosoureas (e.g., carmustine (BCNU) and lomustine (CCNU)), alkylsulphonates (e.g., busulfan and treosulfan), triazenes (e.g., dacarbazine, temozolomide), platinum containing compounds (e.g., cisplatin, carboplatin, oxaliplatin), vinca alkaloids (e.g., vincristine, vinblastine, vindesine, and vinorelbine), taxoids (e.g., paclitaxel or a paclitaxel equivalent such as nanoparticle albumin-bound paclitaxel (ABRAXANE), docosahexaenoic acid bound-paclitaxel (DHA-paclitaxel, Taxoprexin), polyglutamate bound-paclitaxel (PG-paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX), the tumor-activated prodrug (TAP) ANG1005 (Angiopep-2 bound to three molecules of paclitaxel), paclitaxel-EC-1 (paclitaxel bound to the erbB2-recognizing peptide EC-1), and glucose-conjugated paclitaxel, e.g., 2′-paclitaxel methyl 2-glucopyranosyl succinate; docetaxel, taxol), epipodophyllins (e.g., etoposide, etoposide phosphate, teniposide, topotecan, 9-aminocamptothecin, camptoirinotecan, irinotecan, crisnatol, mytomycin C), anti-metabolites, DHFR inhibitors (e.g., methotrexate, dichloromethotrexate, trimetrexate, edatrexate), IMP dehydrogenase inhibitors (e.g., mycophenolic acid, tiazofurin, ribavirin, and EICAR), ribonuclotide reductase inhibitors (e.g., hydroxyurea and deferoxamine), uracil analogs (e.g., 5-fluorouracil (5-FU), floxuridine, doxifluridine, ratitrexed, tegafur-uracil, capecitabine), cytosine analogs (e.g., cytarabine (ara C), cytosine arabinoside, and fludarabine), purine analogs (e.g., mercaptopurine and Thioguanine), Vitamin D3 analogs (e.g., EB 1089, CB 1093, and KH 1060), isoprenylation inhibitors (e.g., lovastatin), dopaminergic neurotoxins (e.g., 1-methyl-4-phenylpyridinium ion), cell cycle inhibitors (e.g., staurosporine), actinomycin (e.g., actinomycin D, dactinomycin), bleomycin (e.g., bleomycin A2, bleomycin B2, peplomycin), anthracycline (e.g., daunorubicin, doxorubicin, pegylated liposomal doxorubicin, idarubicin, epirubicin, pirarubicin, zorubicin, mitoxantrone), MDR inhibitors (e.g., verapamil), Ca.sup.2+ ATPase inhibitors (e.g., thapsigargin), imatinib, thalidomide, lenalidomide, tyrosine kinase inhibitors (e.g., axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTIN™, AZD2171), dasatinib (SPRYCEL®, BMS-354825), erlotinib (TARCEVA®), gefitinib (IRESSA®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib (TYKERB®, TYVERB®), lestaurtinib (CEP-701), neratinib (HKI-272), nilotinib (TASIGNA®), semaxanib (semaxinib, SU5416), sunitinib (SUTENT®, SU11248), toceranib (PALLADIA®), vandetanib (ZACTIMA®, ZD6474), vatalanib (PTK787, PTK/ZK), trastuzumab (HERCEPTIN®), bevacizumab (AVASTIN®), rituximab (RITUXAN®), cetuximab (ERBITUX®), panitumumab (VECTIBIX®), ranibizumab (Lucentis®), nilotinib (TASIGNA®), sorafenib (NEXAVAR®), everolimus (AFINITOR®), alemtuzumab (CAMPATH®), gemtuzumab ozogamicin (MYLOTARG®), temsirolimus (TORISEL®), ENMD-2076, PCI-32765, AC220, dovitinib lactate (TKI258, CHIR-258), BIBW 2992 (TOVOK™), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC1-2036, BMS-690154, CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, and/or XL228), proteasome inhibitors (e.g., bortezomib (VELCADE)), mTOR inhibitors (e.g., rapamycin, temsirolimus (CCI-779), everolimus (RAD-001), ridaforolimus, AP23573 (Ariad), AZD8055 (AstraZeneca), BEZ235 (Novartis), BGT226 (Norvartis), XL765 (Sanofi Aventis), PF-4691502 (Pfizer), GDC0980 (Genetech), SF1126 (Semafoe) and OSI-027 (OSI)), oblimersen, gemcitabine, carminomycin, leucovorin, pemetrexed, cyclophosphamide, dacarbazine, procarbizine, prednisolone, dexamethasone, campathecin, plicamycin, asparaginase, aminopterin, methopterin, porfiromycin, melphalan, leurosidine, leurosine, chlorambucil, trabectedin, procarbazine, discodermolide, carminomycin, aminopterin, and hexamethyl melamine. In certain embodiments, the anti-cancer agent is abiraterone acetate (e.g., ZYTIGA), ABVD, ABVE, ABVE-PC, AC, AC-T, ADE, ado-trastuzumab emtansine (e.g., KADCYLA), afatinib dimaleate (e.g., GILOTRIF), aldesleukin (e.g., PROLEUKIN), alemtuzumab (e.g., CAMPATH), anastrozole (e.g., ARIMIDEX), arsenic trioxide (e.g., TRISENOX), asparaginase erwinia chrysanthemi (e.g., ERWINAZE), axitinib (e.g., INLYTA), azacitidine (e.g., MYLOSAR, VIDAZA), BEACOPP, belinostat (e.g., BELEODAQ), bendamustine hydrochloride (e.g., TREANDA), BEP, bevacizumab (e.g., AVASTIN), bicalutamide (e.g., CASODEX), bleomycin (e.g., BLENOXANE), blinatumomab (e.g., BLINCYTO), bortezomib (e.g., VELCADE), bosutinib (e.g., BOSULIF), brentuximab vedotin (e.g., ADCETRIS), busulfan (e.g., BUSULFEX, MYLERAN), cabazitaxel (e.g., JEVTANA), cabozantinib-s-malate (e.g., COMETRIQ), CAF, capecitabine (e.g., XELODA), CAPOX, carboplatin (e.g., PARAPLAT, PARAPLATIN), carboplatin-taxol, carfilzomib (e.g., KYPROLIS), carmustine (e.g., BECENUM, BICNU, CARMUBRIS), carmustine implant (e.g., GLIADEL WAFER, GLIADEL), ceritinib (e.g., ZYKADIA), cetuximab (e.g., ERBITUX), chlorambucil (e.g., AMBOCHLORIN, AMBOCLORIN, LEUKERAN, LINFOLIZIN), chlorambucil-prednisone, CHOP, cisplatin (e.g., PLATINOL, PLATINOL-AQ), clofarabine (e.g., CLOFAREX, CLOLAR), CMF, COPP, COPP-ABV, crizotinib (e.g., XALKORI), CVP, cyclophosphamide (e.g., CLAFEN, CYTOXAN, NEOSAR), cytarabine (e.g., CYTOSAR-U, TARABINE PFS), dabrafenib (e.g., TAFINLAR), dacarbazine (e.g., DTIC-DOME), dactinomycin (e.g., COSMEGEN), dasatinib (e.g., SPRYCEL), daunorubicin hydrochloride (e.g., CERUBIDINE), decitabine (e.g., DACOGEN), degarelix, denileukin diftitox (e.g., ONTAK), denosumab (e.g., PROLIA, XGEVA), Dinutuximab (e.g., UNITUXIN), docetaxel (e.g., TAXOTERE), doxorubicin hydrochloride (e.g., ADRIAMYCIN PFS, ADRIAMYCIN RDF), doxorubicin hydrochloride liposome (e.g., DOXIL, DOX-SL, EVACET, LIPODOX), enzalutamide (e.g., XTANDI), epirubicin hydrochloride (e.g., ELLENCE), EPOCH, erlotinib hydrochloride (e.g., TARCEVA), etoposide (e.g., TOPOSAR, VEPESID), etoposide phosphate (e.g., ETOPOPHOS), everolimus (e.g., AFINITOR DISPERZ, AFINITOR), exemestane (e.g., AROMASIN), FEC, fludarabine phosphate (e.g., FLUDARA), fluorouracil (e.g., ADRUCIL, EFUDEX, FLUOROPLEX), FOLFIRI, FOLFIRI-BEVACIZUMAB, FOLFIRI-CETUXIMAB, FOLFIRINOX, FOLFOX, FU-LV, fulvestrant (e.g., FASLODEX), gefitinib (e.g., IRESSA), gemcitabine hydrochloride (e.g., GEMZAR), gemcitabine-cisplatin, gemcitabine-oxaliplatin, goserelin acetate (e.g., ZOLADEX), Hyper-CVAD, ibritumomab tiuxetan (e.g., ZEVALIN), ibrutinib (e.g., IMBRUVICA), ICE, idelalisib (e.g., ZYDELIG), ifosfamide (e.g., CYFOS, IFEX, IFOSFAMIDUM), imatinib mesylate (e.g., GLEEVEC), imiquimod (e.g., ALDARA), ipilimumab (e.g., YERVOY), irinotecan hydrochloride (e.g., CAMPTOSAR), ixabepilone (e.g., IXEMPRA), lanreotide acetate (e.g., SOMATULINE DEPOT), lapatinib ditosylate (e.g., TYKERB), lenalidomide (e.g., REVLIMID), lenvatinib (e.g., LENVIMA), letrozole (e.g., FEMARA), leucovorin calcium (e.g., WELLCOVORIN), leuprolide acetate (e.g., LUPRON DEPOT, LUPRON DEPOT-3 MONTH, LUPRON DEPOT-4 MONTH, LUPRON DEPOT-PED, LUPRON, VIADUR), liposomal cytarabine (e.g., DEPOCYT), lomustine (e.g., CEENU), mechlorethamine hydrochloride (e.g., MUSTARGEN), megestrol acetate (e.g., MEGACE), mercaptopurine (e.g., PURINETHOL, PURIXAN), methotrexate (e.g., ABITREXATE, FOLEX PFS, FOLEX, METHOTREXATE LPF, MEXATE, MEXATE-AQ), mitomycin c (e.g., MITOZYTREX, MUTAMYCIN), mitoxantrone hydrochloride, MOPP, nelarabine (e.g., ARRANON), nilotinib (e.g., TASIGNA), nivolumab (e.g., OPDIVO), obinutuzumab (e.g., GAZYVA), OEPA, ofatumumab (e.g., ARZERRA), OFF, olaparib (e.g., LYNPARZA), omacetaxine mepesuccinate (e.g., SYNRIBO), OPPA, oxaliplatin (e.g., ELOXATIN), paclitaxel (e.g., TAXOL), paclitaxel albumin-stabilized nanoparticle formulation (e.g., ABRAXANE), PAD, palbociclib (e.g., IBRANCE), pamidronate disodium (e.g., AREDIA), panitumumab (e.g., VECTIBIX), panobinostat (e.g., FARYDAK), pazopanib hydrochloride (e.g., VOTRIENT), pegaspargase (e.g., ONCASPAR), peginterferon alfa-2b (e.g., PEG-INTRON), peginterferon alfa-2b (e.g., SYLATRON), pembrolizumab (e.g., KEYTRUDA), pemetrexed disodium (e.g., ALIMTA), pertuzumab (e.g., PERJETA), plerixafor (e.g., MOZOBIL), pomalidomide (e.g., POMALYST), ponatinib hydrochloride (e.g., ICLUSIG), pralatrexate (e.g., FOLOTYN), prednisone, procarbazine hydrochloride (e.g., MATULANE), radium 223 dichloride (e.g., XOFIGO), raloxifene hydrochloride (e.g., EVISTA, KEOXIFENE), ramucirumab (e.g., CYRAMZA), R-CHOP, recombinant HPV bivalent vaccine (e.g., CERVARIX), recombinant human papillomavirus (e.g., HPV) nonavalent vaccine (e.g., GARDASIL 9), recombinant human papillomavirus (e.g., HPV) quadrivalent vaccine (e.g., GARDASIL), recombinant interferon alfa-2b (e.g., INTRON A), regorafenib (e.g., STIVARGA), rituximab (e.g., RITUXAN), romidepsin (e.g., ISTODAX), ruxolitinib phosphate (e.g., JAKAFI), siltuximab (e.g., SYLVANT), sipuleucel-t (e.g., PROVENGE), sorafenib tosylate (e.g., NEXAVAR), STANFORD V, sunitinib malate (e.g., SUTENT), TAC, tamoxifen citrate (e.g., NOLVADEX, NOVALDEX), temozolomide (e.g., METHAZOLASTONE, TEMODAR), temsirolimus (e.g., TORISEL), thalidomide (e.g., SYNOVIR, THALOMID), thiotepa, topotecan hydrochloride (e.g., HYCAMTIN), toremifene (e.g., FARESTON), tositumomab and iodine I 131 tositumomab (e.g., BEXXAR), TPF, trametinib (e.g., MEKINIST), trastuzumab (e.g., HERCEPTIN), VAMP, vandetanib (e.g., CAPRELSA), VEIP, vemurafenib (e.g., ZELBORAF), vinblastine sulfate (e.g., VELBAN, VELSAR), vincristine sulfate (e.g., VINCASAR PFS), vincristine sulfate liposome (e.g., MARQIBO), vinorelbine tartrate (e.g., NAVELBINE), vismodegib (e.g., ERIVEDGE), vorinostat (e.g., ZOLINZA), XELIRI, XELOX, ziv-aflibercept (e.g., ZALTRAP), or zoledronic acid (e.g., ZOMETA).
[0332] In certain embodiments, the subject being treated is a mammal. In certain embodiments, the subject is a human. In certain embodiments, the subject is a domesticated animal, such as a dog, cat, cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a companion animal such as a dog or cat. In certain embodiments, the subject is a livestock animal such as a cow, pig, horse, sheep, or goat. In certain embodiments, the subject is a zoo animal. In another embodiment, the subject is a research animal such as a rodent, dog, or non-human primate. In certain embodiments, the subject is a non-human transgenic animal such as a transgenic mouse or transgenic pig.
EXAMPLES
[0333] In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner. Materials and Methods
[0334] All reagents were purchased from commercial suppliers and used without further purification unless stated otherwise. Norbornene anhydride a2.sup.1, dihydroxy oxaliplatin-Pt(IV) al.sup.2, Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat.sup.3, PEG-MM.sup.4, DOX-MM.sup.5, DOX-PC-MM.sup.1, CPT-MM.sup.1, Cy5.5-MM.sup.6, CisPtXL.sup.1, and AcetalXL.sup.5 were prepared according to literature procedures. Liquid chromatography mass spectrometry (LC-MS) tandem was performed on a reverse-phase, C18-column using a binary solvent system (MeCN and H.sub.2O with 0.1% CH.sub.3COOH). Size exclusion chromatography (SEC) analyses were performed on an Agilent 1260 Infinity setup with two Shodex KD-806M columns in tandem and a 0.025 M LiBr DMF mobile phase run at 60° C. The differential refractive index (dRI) of each compound was monitored using a Wyatt Optilab T-rEX detector. Column chromatography was carried out on silica gel 60F (EMD Millipore, 0.040-0.063 mm). Nuclear magnetic resonance (NMR) spectra were recorded on Bruker AVANCE 111-400 spectrometer, with working frequencies of 400 (.sup.1H), 100 (.sup.13C) and 86 (.sup.195Pt) MHz, respectively. Chemical shifts are reported in ppm relative to the signals corresponding to the residual non-deuterated solvents: DMSO-d.sub.6: 5H=2.50 and 5c=39.5 ppm. High-resolution mass spectra (HRMS) were measured on a Bruker Daltonics APEXIV 4.7 Tesla Fourier Transform Ion Cyclotron Resonance Mass Spectrometer (FT-ICR-MS) using an electrospray ionization (ESI) source. Dynamic light scattering (DLS) measurements were performed using a Wyatt Technology Mobius DLS instrument. Samples were prepared by diluting the stock solution (BASP nanoparticles were stored at 0° C. in THF to prevent aggregation) in either nanopure water (MilliQ) or 5% glucose solution (also in nanopure water); the resulting solutions were passed through a 0.4 m Nalgene filter (PES membrane) into disposable polystyrene cuvettes, which were pre-cleaned with compressed air. Measurements were made in sets of 10 acquisitions, and the average hydrodynamic diameters were calculated by fitting the DLS correlation function (Dynamics V7 software package from Wyatt Technology). TEM images were acquired using a FEI Tecnai Multipurpose TEM (G2 Spirit TWIN, 12 kV) at the MIT Center for Materials Science and Engineering. Sample preparation consisted of the following: 5 μL of a 1.0 mg/mL aqueous solution of BASP nanoparticles was pipetted onto a carbon film-coated 200-mesh copper grid (Electron Microscopy Sciences) placed on a piece of parafilm. Next, the aqueous solution was carefully absorbed at the base of the droplet using the edge of a Kimwipe, leaving behind the nanoparticles on the TEM grid. The samples were then stained negatively by adding a drop of 2 wt % uranyl acetate (Electronic Microscopy Sciences). After 5 min, the residual uranyl acetate solution was carefully absorbed onto a Kimwipe, and the samples were allowed to dry completely before analysis.
[0335] RNAi signatures were performed as described previously.sup.2-4. Briefly, Eμ-Myc p19.sup.Arf−/− lymphoma cells were infected at about 30% with GFP-tagged shRNAs. The cells were then treated with drug to kill 80-90% of cells as judged by propidium iodide exclusion via flow cytometry at 48 h. At 72 h, GFP positivity was assessed via flow cytometry. Constrained linear regression was performed in Stata using least-squares fit with individual drug treatments as the predictor variables and the combination signatures as the response variables. The linear regression was constrained such that the sum of the predictor variables had to equal one and none could be negative.
Cell Culture, shRNA Constructs
[0336] Animal Usage:
[0337] All experiments involving animals were reviewed and approved by the MIT Committee for Animal Care (CAC). BALB/c mice (female, 12-16 weeks old) were used for toxicity and pharmacokinetic studies and for therapeutic efficacy study while receiving an alfalfa-free diet (TestDiet) to minimize auto-fluorescence. For statistical significance, all experiments were performed on groups of n=4+. Exclusion criteria included human error in BASP nanoparticle administration.
[0338] In Vivo Toxicity and Pharmacokinetics:
[0339] BASP nanoparticle solutions ranging from 1.0-6.0 mg/200 μL in 5% glucose solution/sterile pH 7.4 PBS buffer were prepared, passed through a sterile 0.2 m filter (Nalgene, PES membrane), and administered into BALB/c mice (n=4) via tail vein injection using a catheter. Adverse physical effects and weight loss were monitored, yielding an optimized dosage of 5.0 mg/200 μL. For pharmacokinetic study, BASP nanoparticle doses (4.5 mg/200 μL) were injected into BALB/c mice (3.0 groups of n=3), and blood samples were taken at 0, 0.25, 5, 24, and 48 h via cheek bleed. Samples were centrifuged to obtain blood serum, which were then subjected to fluorescence imaging (IVIS, Cy5.5 λ.sub.ex/λ.sub.em=640/700 nm, Xenogen). Additionally, tumor localization was monitored at 3 and 24 h, followed by euthanizing the mice and subjecting the excised organs to fluorescence imaging.
[0340] In Vivo Therapeutic Efficacy:
[0341] Ovarian cancer cells (SKOV-3; ATCC) were grown in RPMI-1640 media supplemented with 0.01 mg/mL bovine insulin, 20% fetal bovine serum in 5% CO.sub.2 humidified atmosphere (37° C.) to a final concentration of 20%; cells were then harvested, mixed with Matrigel and sterile pH 7.4 PBS buffer, filtered through sterile 0.2 m filters, and subcutaneously injected on the flanks of the mice. Tumor growth was monitored until 0.9 cm in cumulative diameter was reached. At this point, drug-loaded BASP nanoparticles were injected (5.0 mg/200 μL, weekly injections) to the treatment group (n=4) while the same amount of 5% glucose were administered to the control group (n=4) via tail vein injection. Tumor progression and therapeutic efficacy were monitored via IVIS imaging, caliber and/or ImageJ measurements. At set time-points, mice were taken out of the study for blood chemistry panel analysis, tumor and/or organ collection, followed by histology/pathology analysis.
[0342] In Vivo IVIS Imaging:
[0343] In vivo imaging was performed on an IVIS Spectrum-bioluminescent and fluorescent imaging system (Xenogen) at the Koch Institute for Integrative Cancer Research at MIT. Mice were anesthetized in isoflurane chamber for 10 min prior to imaging. Fluorescence imaging was acquired based on the Cy5.5 component of the BASP nanoparticles (λ.sub.ex/λ.sub.em=675/nm, exposure time 2-10 s).
Example 1: Oxaliplatin Crosslinker (OxPtXL): Synthetic Protocol
[0344] Oxaliplatin Crosslinker (OxPtXL): A modified procedure was used in the synthesis of OxPtXL (
Example 2: Procedure for Brush-Arm Star Polymer (BASP) Syntheses
[0345] All BASP syntheses were performed in a glovebox under a N.sub.2 atmosphere; however, similar results are expected under ambient conditions. All ROMP reactions followed the same general procedure, which was modified from previously published literature.sup.1,5
[0346] Chemical Structures of all Macromonomers (MMs) and Crosslinkers (XLs):
[0347] The chemical structures of macromonomers (MMs) and crosslinkers (XLs) used are shown in
[0348] General Scheme for Brush Polymerization and Crosslinking to Form BASPs:
[0349] The general scheme for the synthesis of the reported drug-loaded brush, followed by cross-linking with the bis-norbornene functionalized Pt(IV) prodrug to afford brush-arm star polymers (BASPs) is shown in
[0350] Cisplatin(cisPt)-Doxorubicin(DOX)-Camptothecin(CPT) Nanoparticles:
[0351] The general scheme for the synthesis of cisPt-DOX-CPT nanoparticles is shown in
[0352] DOX-Loaded BASP (NP-DOX):
[0353] To a 4 mL vial, a suspension of AcetalXL (5.70 mg, 9.80 μmol, 15.0 eq) in THF (98.0 μL, 0.1 M AcetalXL) was prepared. To a second 4 mL vial containing a stir bar, DOX-MM (3.80 mg, 0.91 μmol, 1.4 eq) and PEG-MM (20.00 mg, 5.63 μmol, 8.6 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (33.0 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (32.7 μL, 0.65 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 10:1, while achieving a total MM concentration of 0.1M, affording a red solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The AcetalXL suspension was then added into the MM vial in three aliquots (5 equiv. each addition) until the desired 15.0 equiv. were added, and the polymerizing mixture was allowed to stir for 1 h at room temperature, affording a red solution. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. (
[0354] OxPt-Loaded BASP (NP-OxPt):
[0355] To a 4 mL vial containing a stir bar, OxPtXL (3.82 mg, 4.57 μmol, 5.0 eq) was added. To a second 4 mL vial containing a stir bar, PEG-MM (30.00 mg, 9.13 μmol, 10.0 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (136.9 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (45.6 μL, 0.91 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 10:1, while achieving a total MM concentration of 0.05M, affording a yellow solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The polymerizing mixture was then transferred (163 μL) to the XL vial, and the solution was allowed to stir for 6 h at room temperature. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. (
[0356] PEG BASP (NP-PEG):
[0357] To a 4 mL vial, a suspension of AcetalXL (23.50 mg, 40.5 μmol, 14.3 eq) in THF (405 μL, 0.1 M AcetalXL) was prepared. To a second 4 mL vial containing a stir bar, PEG-MM (65.00 mg, 19.80 μmol, 7.0 eq) was added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (100 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (98.0 μL, 2.82 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 7:1, while achieving a total MM concentration of 0.1M, affording a yellow solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The AcetalXL suspension was then added dropwise (in 3 aliquots of ˜130 μL) over the course of 10 min into the MM vial, and the polymerizing mixture was allowed to stir for 3 h at room temperature, affording a brown solution. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. (
[0358] CPT-DOX-CisPt BASP (NP-3D-CisPt):
[0359] To a 4 mL vial containing a stir bar, CisPtXL (0.82 mg, 1.12 μmol, 3.0 eq) was added. To a second 4 mL vial containing a stir bar, CPT-MM (3.01 mg, 0.77 μmol, 2.1 eq), DOX-MM (1.27 mg, 0.31 μmol, 0.8 eq), and PEG-MM (5.00 mg, 1.52 μmol, 4.1 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (33.4 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (18.6 μL, 0.37 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 7:1, while achieving a total MM concentration of 0.05M, affording a red solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The polymerizing mixture was then transferred to the XL vial, and the solution was allowed to stir for 6 h at room temperature. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C.
[0360] CPT-DOX-PC-OxPt BASP (NP-3D-PC-OxPt):
[0361] To a 4 mL vial containing a stir bar, OxPtXL (7.42 mg, 8.86 μmol, 5.0 eq) was added. To a second 4 mL vial containing a stir bar, CPT-MM (24.00 mg, 6.13 μmol, 3.5 eq), DOX-PC-MM (10.27 mg, 2.46 μmol, 1.4 eq), and PEG-MM (30.00 mg, 9.13 μmol, 5.2 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (266 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (88.4 μL, 1.77 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 10:1, while achieving a total MM concentration of 0.05M, affording a red solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The polymerizing mixture was then transferred (334 μL) to the XL vial, and the solution was allowed to stir for 6 h at room temperature. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. The D.sub.H for the NP-3D-PC-OxPt BASP nanoparticles was determined to be 16±3 nM.
[0362] CPT-Diluted DOX-PC-OxPt BASP (NP-3D-DD-OxPt):
[0363] To a 4 mL vial containing a stir bar, OxPtXL (7.66 mg, 9.14 μmol, 5.0 eq) was added. To a second 4 mL vial containing a stir bar, CPT-MM (35.77 mg, 9.14 μmol, 5.0 eq), DOX-PC-MM (0.0763 mg, 0.0183 μmol, 0.01 eq), and PEG-MM (30.00 mg, 9.13 μmol, 4.99 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (274 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (91.9 μL, 1.83 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 10:1, while achieving a total MM concentration of 0.05M, affording a yellow solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The polymerizing mixture was then transferred (346 L) to the XL vial, and the solution was allowed to stir for 6 h at room temperature. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. The D.sub.TEM for the NP-3D-DD-OxPt BASP nanoparticles was determined to be 57±33 nM and the D.sub.H was determined to be 37±21 nM.
[0364] CPT-DOX-OxPt BASP (NP-3D-OxPt):
[0365] To a 4 mL vial containing a stir bar, OxPtXL (7.63 mg, 9.11 μmol, 5.0 eq) was added. To a second 4 mL vial containing a stir bar, CPT-MM (24.66 mg, 6.30 μmol, 3.5 eq), DOX-MM (10.59 mg, 2.53 μmol, 1.4 eq), and PEG-MM (30.00 mg, 9.38 μmol, 5.2 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (273 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (91.0 μL, 1.82 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 10:1, while achieving a total MM concentration of 0.05M, affording a red solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The polymerizing mixture was then transferred (344 μL) to the XL vial, and the solution was allowed to stir for 6 h at room temperature. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. (
[0366] CPT-DOX-OxPt-Cy5.5 BASP (NP-3D-OxPt-Cy5.5):
[0367] To a 4 mL vial containing a stir bar, OxPtXL (7.82 mg, 9.34 μmol, 5.0 eq) was added. To a second 4 mL vial containing a stir bar, CPT-MM (25.30 mg, 6.46 μmol, 3.5 eq), DOX-MM (10.86 mg, 2.60 μmol, 1.4 eq), Cy5.5-MM (1.00 mg, 0.25 μmol, 0.13 eq), and PEG-MM (30.00 mg, 9.38 μmol, 5.0 eq) were added. To a third vial, a solution of Grubbs 3.sup.rd generation bispyridyl catalyst G3-Cat (0.02 M in THF) was freshly prepared. THF (275 μL) was then added to the MM vial, followed by the addition of the G3-Cat solution (93.4 μL, 1.87 μmol, 1.0 eq) to give the desired MM:G3-Cat ratio of 10:1, while achieving a total MM concentration of 0.05M, affording a dark brown solution. The reaction mixture was allowed to stir for 20 min at room temperature before an aliquot (˜10 μL) was taken out and quenched with 1 drop of ethyl vinyl ether for GPC analysis. The polymerizing mixture was then transferred to the XL vial, and the solution was allowed to stir for 6 h at room temperature. To quench the polymerization, a drop of ethyl vinyl ether was then added. The newly formed nanoparticles were transferred to an 8 kD MW cutoff dialysis tubing (Spectrum Laboratories) in 5 mL THF, and the solution was dialyzed against THF (500 mL×3, solvent exchange every 6 h). The BASP nanoparticles were stored in THF at 0° C. (
[0368] Free Drug Combination (DOX-CPT-OxPt): A stock solution of free DOX and CPT at 0.50 mg/mL (DOX) and 0.79 mg/mL (CPT) in 5% glucose was prepared. To a second vial, a free OxPt stock solution (24.75 mg/mL) in DMSO was prepared. The OxPt solution was diluted 20-fold with the DOX-CPT solution, forming the final free drug solution with 0.47 mg/mL DOX, 0.75 mg/mL CPT, and 1.24 mg/mL OxPt. The final solution contained 5% DMSO; the net amount and composition of drug delivered was matched to the BASP system at 200 μL injections.
[0369] General Scheme for Iterative Exponential Growth+Analysis:
[0370] A general scheme analysis for the IEG+ is shown in
[0371] Analysis of Homopolymer Blocks:
[0372] A general synthesis of homopolymer blocks was performed, resulting in a 32-unit polymer (
[0373] Analysis of Alternating Copolymer Blocks:
[0374] A general synthesis of alternating copolymer blocks was performed with further analysis done using MALDI and GPC (
Example 3: RNAi Signature Assay
[0375] Heat Map and Principle Component Analysis of UV-Stimulated and Diluted DOX NPs:
[0376] A signature assay heat map (
[0377] RNAi Signatures and Classification of Individual Free Drugs, Prodrugs, and Nanoparticles:
[0378] Eμ-Myc p19.sub.Ar−/− lymphoma cells were infected at about 30% with GFP-tagged shRNAs. The cells were then treated with drug to kill 80-90% of cells as judged by propidium iodide exclusion via flow cytometry at 48 h. At 72 h GFP positivity was assessed via flow cytometry. Using the RNAi signature approach, the free drug, prodrug, single-drug-conjugated BASPs, and multi-drug conjugated BASPs illustrated in
[0379] Contribution of Different Drugs to Mechanism of Action of Combination Nanoparticles as Predicted by Constrained Linear Regression:
[0380] Constrained linear regression was performed in Matlab 2015a using the ‘Isqlin’ function with individual drug treatments as the predictor variables and the combination signatures as the response variables. The linear regression was constrained such that the sum of the predictor variables had to equal one and none could be negative. This was repeated for all combinations of replicates for both the response and predictor variables. This resulted in a minimum of 120 individual linear regressions. The results of all regressions were then averaged to obtain the final result. Error is the standard error of the mean of all of the regressions. In order to use RNAi-based signatures to determine the relative effective contribution of multiple drugs within a single NP, a BASP capable of externally triggered drug release was designed to easily validate the presence or absence of the triggered drug in a combination of other drugs without having to change the BASP composition. For this, CPT-MM and photocleavable DOX-PC-MM were crosslinked with OxPtXL via BF-ROMP to yield 3-drug-conjugated BASP NP-3D-PC-OxPt. The incorporation of the UV-triggered DOX-PC-MM allows for rapid release of free DOX upon exposure to 365 nm light. NP-3D-PC-OxPt was used to treat cells either with or without light irradiation and RNAi signatures were obtained. Constrained linear regression was then performed (
Example 4: Histology
[0381] Comparison of H&E Stained Liver Cross Sections of Treated and Untreated Mice:
[0382] Different views of paraffin-embedded and H&E stained liver cross sections were obtained from control mice (
[0383] Comparison of H&E Stained Tumor Cross Sections of Treated and Untreated Mice:
[0384] Different views of paraffin-embedded and H&E stained left (L) and right (R) tumor cross sections were obtained from control mice (
[0385] Comparison of H&E Stained Tumor and Liver Cross Sections of Treated and Untreated Mice:
[0386] Different views of H&E stained left (L) and right (R) tumor cross sections obtained from a control mouse (
Example 5: Blood Chemistry Analysis
[0387] Pharmacokinetics of OxPt-Loaded BASP (NP-OxPt):
[0388] Pharmacokinetics over a 126 h period associated with NP-OxPt containing 1% Cy5.5-MM injected (4.5 mg/dose; 18 mg/kg) in BALB/c mice are shown in
[0389] Blood Panel Analysis of Nanoparticle/Free-Drug Treated and Control NCR-NU Mice:
[0390] Blood panel analysis of biomarkers associated with NCR-NU mice treated with: i) NP-3D-OxPt (#114 bars: Chronic; #19 bars: Acute); ii) the free drug combination (#5 bars: DOX-CPT-OxPt); and iii) 5% glucose solutions is shown in
Example 6: In Vivo Efficacy
[0391] Animal Usage:
[0392] All experiments involving animals were reviewed and approved by the MIT Committee for Animal Care (CAC). BALB/c mice (female, 8-12 weeks old, Taconic) were used for toxicity and pharmacokinetic studies. NCR-NU nude mice (female, 8-12 weeks old, Taconic) were used for biodistribution and therapeutic efficacy studies while receiving an alfalfa-free diet (TestDiet) to minimize auto-fluorescence. For statistical significance, all experiments were performed on groups of n=4+.
[0393] Tumor Volume Plot, Survival Curve, and Nanoparticle Localization in 1 cm Diameter Tumor Study:
[0394] Ovarian cancer cells (SKOV-3; ATCC) were grown in RPMI-1640 media supplemented with 0.01 mg/mL bovine insulin, 20% fetal bovine serum in 5% CO.sub.2 humidified atmosphere (37° C.) to a final concentration of 20%. The cells were then harvested, mixed with Matrigel and sterile pH 7.4 PBS buffer (1:1), filtered through sterile 0.2 μm filters, and injected subcutaneously (1.25×106 cells) in the hind flanks of the mice. Tumor growth was monitored for 2-4 wks until either 0.5 or 1.0 cm in cumulative diameter was reached as the starting point for the two efficacy studies, after which treatment groups were randomized. At this point, three-drug-loaded BASP nanoparticles (NP-3D-OxPt) were injected (5.0 mg/200 μL, approx. weekly injections) to the treatment group (n=4+), while the same volume of 5% glucose was administered to the control group (n=4+) via tail vein injection. Tumor progression and therapeutic efficacy were monitored via caliper and ImageJ measurements. At set time-points, mice were removed from the study for blood chemistry panel analysis (Charles River), tumor and/or organ excision, followed by histology and pathology analysis. Non-sacrificial mice were removed from the study if one or more of the following criteria were met: i) body weight decreased by 10% or more compared to the body weight at the start of the study, ii) the diameter of the tumor doubled compared to the starting diameter (i.e., if starting with 0.5 cm diameter, then the mouse was removed at 1.0 cm; or if starting at 1.0 cm, then the mouse was removed at 2.0 cm in diameter), or iii) development of necrotic, and/or cracked and bleeding tumor tissue. As discussed above, tumor localization of NP-3D-OxPt-Cy5.5 was investigated by injecting NCR-NU mice with 5 mg of the three-drug-loaded nanoparticle and monitoring (excitation at 675 nm; emission at 720 nm) the epifluorescence at 3 and 20 h in an IVIS whole animal imaging system (
[0395] Tumor Volume Plot, Survival Curve, and Nanoparticle Localization in 0.5 cm Diameter Tumor Study:
[0396] To investigate the therapeutic efficacy of the three-drug-loaded NP-3D-OxPt in vivo, NCR-NU mice were injected subcutaneously in each hind flank with 1.25×106 ovarian carcinoma cells (SKOV-3, ATCC) mixed (1:1) with Matrigel and PBS buffer. Tumor growth was monitored for 2-4 wks until the tumor reached approximately 0.5 cm in diameter, at which point three treatment groups were established: those treated with sterile-filtered 5% aqueous glucose solutions containing either i) NP-3D-OxPt, ii) a free drug formulation at the same DOX, CPT, and OxPt concentrations as NP-3D-OxPt, or iii) the blank vehicle. The BASP treatment schedule consisted of four tail-vein injections over 22 days (˜1 inj./wk), where each injection per mouse comprised of 5 mg BASP in 200 μL of 5% glucose solution, a dose that is close to the maximum solubility limit (˜6 mg/200 μL) of the three-drug-conjugated BASP. Since ˜10% of each NP-3D-OxPt is made up of the three anti-cancer drugs, each dose is equal to ˜20 mg total drug/kg mouse. Although the ratio of drug loading in NP-3D-OxPt should afford maximum tolerable doses (MTD) that match those of free DOX and CPT, and more than twice that of OxPt, the results from the RNAi signature assay suggest that, at least in vitro, the effective contribution from all three drugs toward the overall mechanism of action should be nearly equal. Simultaneous delivery of all three drugs within one BASP entity ensures that the three drugs will arrive at the tumor in a ratio defined by the BASP, which precludes differences in pharmacokinetics and biodistribution that could be observed for mixtures of single-drug-conjugated NPs. Thus, results obtained at the cell culture level can be expected to translate to the tumor. As a consequence of the branched MM design, the BASP possesses a protective PEG outer layer, a feature that greatly enhances its bioavailability and leads to a blood circulation half-life (t1/2) of 43±8 h for NP-OxPt in NCR-NU mice (
[0397] Tumor Reduction:
[0398] Nearly complete tumor reduction was observed in Mouse #105 over a 21 day treatment schedule and 12 week observation using CPT-DOX-OxPt (
DISCUSSION
[0399] It has been demonstrated that a modular BASP combination therapy platform—comprised of drug-conjugated MMs and Pt(IV)-based XLs—is an effective means of delivering a precise ratio of an otherwise toxic combination of three drugs to a subcutaneous xenograft tumor in mice. Previously, there was no way to verify that the drug mechanism of action was not disrupted by incorporation into the nanoparticle. The mechanism of action associated with the reported prodrugs and single-drug-loaded BASPs was characterized using an RNAi signature assay that allows for a fast and accurate in vitro combinatorial screening that is capable of predicting a prodrug's mechanism of action. Although the Pt(IV)-diester XL—intended to be a precursor to CisPt—did not behave as anticipated in vitro, the RNAi signatures allowed the identification of another Pt(IV)-diester XL that functions as a faithful precursor to OxPt. Thus, a functional genetic assay was employed to ensure that the conjugated drugs—whose mechanistic targets differ from one another—behaved as intended. Moreover, constrained linear regression analysis, paired with RNAi signatures, revealed the relative effective contributions of each drug towards the mechanism of action in our three-drug-conjugated BASPs. The ability to assess the in vitro contribution of each drug adds another layer of quality control to NP-based drug delivery. The initial stoichiometric ratios can be tuned to achieve BASPs where all drugs contribute equally, or BASPs that possess disparate drug contributions that may potentially maximize synergistic ratios, all the while taking into account the kinetics associated with the release of each drug from the combination NP platformThis modular platform and the RNAi-based mechanism of action predictive assay can be further utilized to assess different combinations of drugs in an effort to treat other types of cancer, as well as characterize nanoparticles other than BASPs consisting of one, two, three, or potentially more drugs.
REFERENCES
[0400] .sup.1 Liao, L.; Liu, J.; Dreaden, E. C.; Morton, S. W.; Shopsowitz, K. E.; Hammond, P. T.; Johnson, J. A. J. Am. Chem. Soc. 2014, 136, 5896-5899. [0401] .sup.2 Hall, M.; Dillon, C.; Zhang, M.; Beale, P.; Cai, Z.; Lai, B.; Stampfl, A. J.; Hambley, T. J. Biol. Inorg. Chem. 2003, 8, 726-732. [0402] .sup.3 Love, J. A.; Morgan, J. P.; Trnka, T. M.; Grubbs, R. H. Angew. Chem. Int. Ed. 2002, 41, 4035-4037. [0403] .sup.4 Liu, J.; Burts, A. O.; Zhukhovitsky, A. V.; Ottaviani, M. F.; Turro, N. J.; Johnson, J. A. J. Am. Chem. Soc. 2012, 134, 16337-16344 [0404] .sup.5 Gao, A. X.; Liao, L.; Johnson, J. A. ACS Macro Lett. 2014, 3, 854-857. [0405] .sup.6 Sowers, M. A; McCombs, J. R.; Wang, Y.; Paletta, J. T.; Morton, S. W.; Dreaden, E. C.; Boska, M. D.; Ottaviani, M. F.; Hammond, P. T.; Rajca, A.; Johnson, J. A. Nature Commun. 2014, 5, 5460.
EQUIVALENTS AND SCOPE
[0406] In the claims articles such as “a,” “an,” and “the” may mean one or more than one unless indicated to the contrary or otherwise evident from the context. Claims or descriptions that include “or” between one or more members of a group are considered satisfied if one, more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process unless indicated to the contrary or otherwise evident from the context. The invention includes embodiments in which exactly one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.
[0407] Furthermore, the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, and descriptive terms from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim. Where elements are presented as lists, e.g., in Markush group format, each subgroup of the elements is also disclosed, and any element(s) can be removed from the group. It should it be understood that, in general, where the invention, or aspects of the invention, is/are referred to as comprising particular elements and/or features, certain embodiments of the invention or aspects of the invention consist, or consist essentially of, such elements and/or features. For purposes of simplicity, those embodiments have not been specifically set forth in haec verba herein. It is also noted that the terms “comprising” and “containing” are intended to be open and permits the inclusion of additional elements or steps. It is also noted that the term “including” is intended to be open and permits the inclusion of additional elements or steps. Where ranges are given, endpoints are included. Furthermore, unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.
[0408] This application refers to various issued patents, published patent applications, journal articles, and other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the invention can be excluded from any claim, for any reason, whether or not related to the existence of prior art.
[0409] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments described herein. The scope of the present embodiments described herein is not intended to be limited to the above Description, but rather is as set forth in the appended claims. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.