Platinum-containing compounds, and related compositions and uses thereof
11203608 · 2021-12-21
Assignee
Inventors
- Stephen J. Lippard (Washington, DC, US)
- Omer Yilmaz (Cambridge, MA, US)
- Fang Wang (Cambridge, MA, US)
- Jonathan Braverman (Cambridge, MA, US)
Cpc classification
A61P35/00
HUMAN NECESSITIES
International classification
Abstract
Platinum-containing compounds are generally described. For example, compounds of Formula (II) are generally described. Inventive compositions and uses thereof are also described. For example, methods of treating cancer in a subject comprising administering to the subject a therapeutically effective amount of a compound of Formula (II) are generally described. ##STR00001##
Claims
1. A compound of Formula (II): ##STR00020## or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof; wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.6, and R.sup.7 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM; wherein M is a cation; wherein R.sup.5 and R.sup.8 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM; wherein R.sup.9 and R.sup.12 are each independently selected from the group consisting of —CR(R′)—, carbonyl, imine, alkylimine, and arylimine; wherein R and R′ are independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, and diarylamino; wherein R.sup.10 and R.sup.11 are each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, —(C═O)—(CH.sub.2).sub.kR.sup.15, —(CHOR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(C═NR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(CHNHR.sup.16)—(CH.sub.2).sub.kR.sup.15, and —(CHNR.sup.16.sub.2)—(CH.sub.2).sub.kR.sup.15; wherein R.sup.15 is selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, and siloxy; wherein R.sup.16 is selected from the group consisting of hydrogen, alkyl, aryl, and silyl; wherein k is 0, 1, 2, or 3; wherein R.sup.13 and R.sup.14 are each independently selected from the group consisting of hydrogen, halogen, alkyl, and aryl; wherein A.sup.1 and A.sup.2 are each independently selected from the group consisting of oxygen, sulfur, and —NR.sup.17—; wherein R.sup.17 is selected from the group consisting of hydrogen, alkyl, and aryl; wherein X.sup.1 and X.sup.2 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, oxygen, hydroxy, alkoxy, aryloxy, siloxy, sulfur, thiol, alkyl sulfide, aryl sulfide, alkyl sulfoxide, aryl sulfoxide, sulfinate, selenium, selenol, alkyl selenide, aryl selenide, alkyl selenoxide, aryl selenoxide, and seleninate; wherein X.sup.3 and X.sup.4 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, heteroarylene, water, halide, carboxylate, hydroxide, alkoxide, aryloxide, siloxide, dialkyl sulfide, diaryl sulfide, alkyl aryl sulfide, dialkyl sulfoxide, diaryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfinate, aryl sulfinate, alkyl sulfonate, aryl sulfonate, sulfite, sulfate, thiosulfate, dialkyl selenide, diaryl selenide, alkyl aryl selenide, dialkyl selenoxide, diaryl selenoxide, alkyl aryl selenoxide, selenite, and seleninate; wherein Y and Z are each independently selected from the group consisting of hydroxide, alkoxide, aryloxide, siloxide, and halide; wherein n is 0 or 1; wherein m is 0 or 1; and wherein M, Y, Z, A.sup.1, A.sup.2, X.sup.1, X.sup.2, X.sup.3, X.sup.4, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and/or R.sup.17 are each independently optionally substituted.
2. The compound of claim 1, wherein X.sup.1 and/or X.sup.2 is amino.
3. The compound of claim 1, wherein R.sup.2, R.sup.3, R.sup.4, and/or R.sup.13 are hydrogen.
4. The compound of claim 1, wherein R.sup.6, R.sup.7, and/or R.sup.10 are hydroxy, and R.sup.9 and/or R.sup.12 are methylene.
5. The compound of claim 1, wherein A.sup.1, A.sup.2, R.sup.5 and/or R.sup.8 are oxygen, and R.sup.1 is alkoxy.
6. The compound of claim 1, wherein R.sup.11 is —(C═O)—(CH.sub.2).sub.kR.sup.15, R.sup.15 is hydroxyl, and k is 1.
7. The compound of claim 1, wherein n and m are both 0.
8. A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the compound of claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.
9. The method of claim 8, wherein the cancer is selected from the group consisting of ovarian cancer, colorectal cancer, breast cancer, lung cancer, prostate cancer, osteosarcoma, and/or leukemia.
10. The method of claim 8, wherein the subject is a human.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1)
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10)
(11)
(12)
DETAILED DESCRIPTION
(13) Compounds, compositions thereof, preparative methods thereof, and related therapeutic methods are disclosed herein. For example, disclosed herein are platinum-containing compounds, and compositions and uses thereof. In some embodiments, the platinum-containing compound comprises a moiety that comprises a portion of an anthracycline drug (e.g., a moiety that comprises a portion of doxorubicin) conjugated to a platinum-containing moiety. In certain embodiments, the compounds disclosed herein, and related compositions thereof, can be used for treating a disease in a subject in need thereof. For example, in some cases, the compounds disclosed herein, and related compositions thereof, can be used for treating cancer (e.g., ovarian cancer, colorectal cancer, breast cancer, lung cancer, prostate cancer, osteosarcoma, and/or leukemia), as well as other diseases caused by abnormal cell proliferation and growth in a subject in need thereof.
(14) As discussed above, there is increasing resistance to platinum-based agents and to anthracycline drugs. The in vivo efficacy of two agents (e.g., a platinum-based agent and an anthracycline drug) when merely combined is frequently significantly attenuated. This attenuation can be due, in part, to the different pharmacokinetics and tissue distributions of each drug, which can lead to difficulty in delivering them to the target at the respective optimal concentrations. However, it was unexpectedly discovered that, in some embodiments, the chemical conjugation of platinum-based agents to anthracyclines through non-labile covalent bonds modulates the activity and toxicity profile of both agents. In some embodiments, the two therapeutically active components are able to reach their destination molecular target together in cancer cells and simultaneously exert their cytotoxic activities. Thus, it was unexpectedly discovered that the product of covalently conjugating two pharmacophores might, in certain embodiments, be superior to the simple physical combination of the same two agents. In certain embodiments, a platinum-anthracycline conjugate might exhibit additional beneficial anticancer properties. For example, in some cases, the conjugate might act against cancer cells by the mechanisms of both the platinum agent and the anthracycline, thereby increasing its potency relative to either drug alone. In certain instances, cells resistant to the platinum agent, the anthracycline, or both might still be sensitive to the platinum-anthracycline conjugate. Additional details regarding the platinum-containing compounds, and compositions and uses thereof, are provided below.
(15) Certain aspects are related to platinum-containing compounds.
(16) In some embodiments, the compound is a compound of Formula (I):
(17) ##STR00004##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof;
(18) wherein X.sup.1 and X.sup.2 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, oxygen hydroxy, alkoxy, aryloxy, siloxy, sulfur, thiol, alkyl sulfide, aryl sulfide, alkyl sulfoxide, aryl sulfoxide, sulfinate, selenium, selenol, alkyl selenide, aryl selenide, alkyl selenoxide, aryl selenoxide, and seleninate;
(19) wherein X.sup.3 and X.sup.4 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, heteroarylene, water, halide, carboxylate, hydroxide, alkoxide, aryloxide, siloxide, dialkyl sulfide, diaryl sulfide, alkyl aryl sulfide, dialkyl sulfoxide, diaryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfinate, aryl sulfinate, alkyl sulfonate, aryl sulfonate, sulfite, sulfate, thiosulfate, dialkyl selenide, diaryl selenide, alkyl aryl selenide, dialkyl selenoxide, diaryl selenoxide, alkyl aryl selenoxide, selenite, and seleninate;
(20) wherein Y and Z are each independently selected from the group consisting of hydroxide, alkoxide, aryloxide, siloxide, and halide;
(21) wherein n is 0 or 1;
(22) wherein m is 0 or 1; and
(23) wherein A, Y, Z, X.sup.1, X.sup.2, X.sup.3, and/or X.sup.4 are each independently optionally substituted.
(24) In certain embodiments, A comprises at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of the structure of an anthracycline drug. In some instances, A comprises less than or equal to 100%, less than or equal to 99%, less than or equal to 95%, less than or equal to 90%, or less than or equal to 80% of the structure of an anthracycline drug. Combinations of these ranges are also possible (e.g., 90-95% of the structure of an anthracycline drug).
(25) The percentage of the anthracycline drug included in A is calculated by determining the percentage of the molecular weight of the portions of the anthracycline drug included in A versus the molecular weight of the anthracycline drug. For example, if the compound comprises doxaliplatin—a compound of Formula (I)—(shown below on the left), wherein A is the portion of doxaliplatin that is not in the box, and wherein A differs from doxorubicin (shown below on the right) only in that the boxed portion of doxorubicin is not included in A (because the two amino groups in the box shown in doxaliplatin are X.sup.1 and X.sup.2 of Formula (I) rather than part of A), A comprises 93.92% of the structure of an anthracycline drug (doxorubicin):
(26) ##STR00005##
If the molecular weight of doxorubicin is 543.52 g/mol, and A includes all of doxorubicin except an NH.sub.2 group and an OH group, which have molecular weights of 16.02 g/mol and 17.00 g/mol, respectively, then A comprises 93.92% ((543.52-16.02-17.00)/543.52×100) of the anthracycline drug.
(27) In accordance with some embodiments, the portion of the anthracycline drug included in A is at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% of A. In certain embodiments, the portion of the anthracycline drug included in A is less than or equal to 100%, less than or equal to 99%, less than or equal to 95%, less than or equal to 90%, or less than or equal to 80% of A. Combinations of these ranges are also possible (e.g., 90-99% of A is the portion of the anthracycline drug).
(28) The percentage of A that is the portion of the anthracycline drug is calculated by determining the percentage of the molecular weight of the portion of the anthracycline drug versus the total molecular weight of A. In the example discussed above, the molecular weight of the portion of the anthracycline drug (doxorubicin) included in A was 510.5 g/mol (543.52-16.02-17.00), and the total molecular weight of A was 510.5 g/mol; thus, in this example, the portion of the anthracycline drug included in A is 100% of A.
(29) In some embodiments, the anthracycline drug is FDA-approved. In certain embodiments, the anthracycline drug is not FDA-approved. In some cases, the anthracycline drug is selected from the group consisting of doxorubicin, epirubicin, daunorubicin, idarubicin, and valrubicin. For example, in certain instances, the anthracycline drug is doxorubicin.
(30) In some instances, X.sup.1 and/or X.sup.2 are an amino group. For example, in doxaliplatin, both X.sup.1 and X.sup.2 are NH.sub.2.
(31) In certain embodiments, X.sup.3 and/or X.sup.4 is a halide. In accordance with some embodiments, X.sup.3 and/or X.sup.4 is a carboxylate. For example, in doxaliplatin, both X.sup.3 and X.sup.4 are a carboxylate. X.sup.3 and X.sup.4 are joined together to form a bidentate ligand, in certain cases. For example, in doxaliplatin, X.sup.3 and X.sup.4 are joined together to form a bidentate ligand. According to some embodiments, X.sup.3 and X.sup.4 can be joined with one of X.sup.1 and X.sup.2 to form a tridentate ligand. Still further, in certain embodiments, both X.sup.3 and X.sup.4 can be joined with X.sup.1 and X.sup.2 to form a tetradentate ligand.
(32) In some cases, n and m are both 1. In other cases, n and m are both 0. For example, in doxaliplatin, n and m are both 0.
(33) In some embodiments, the compound is a compound of Formula (II):
(34) ##STR00006##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof;
(35) wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.6, and R.sup.7 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM;
(36) wherein M is a cation;
(37) wherein R.sup.5 and R.sup.8 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM;
(38) wherein R.sup.9 and R.sup.12 are each independently selected from the group consisting of —CR(R′)—, carbonyl, imine, alkylimine, and arylimine;
(39) wherein R and R′ are independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, and diarylamino;
(40) wherein R.sup.10 and R.sup.11 are each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, —(C═O)—(CH.sub.2).sub.kR.sup.15, —(CHOR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(C═NR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(CHNHR.sup.16)—(CH.sub.2).sub.kR.sup.15, and —(CHNR.sup.16.sub.2)—(CH.sub.2).sub.kR.sup.15;
(41) wherein R.sup.15 is selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, and siloxy;
(42) wherein R.sup.16 is selected from the group consisting of hydrogen, alkyl, aryl, and silyl;
(43) wherein k is 0, 1, 2, or 3;
(44) wherein R.sup.13 and R.sup.14 are each independently selected from the group consisting of hydrogen, halogen, alkyl, and aryl;
(45) wherein A.sup.1 and A.sup.2 are each independently selected from the group consisting of oxygen, sulfur, and —NR.sup.17;
(46) wherein R.sup.17 is selected from the group consisting of hydrogen, alkyl, and aryl;
(47) wherein X.sup.1 and X.sup.2 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, oxygen, hydroxy, alkoxy, aryloxy, siloxy, sulfur, thiol, alkyl sulfide, aryl sulfide, alkyl sulfoxide, aryl sulfoxide, sulfinate, selenium, selenol, alkyl selenide, aryl selenide, alkyl selenoxide, aryl selenoxide, and seleninate; wherein X.sup.3 and X.sup.4 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, heteroarylene, water, halide, carboxylate, hydroxide, alkoxide, aryloxide, siloxide, dialkyl sulfide, diaryl sulfide, alkyl aryl sulfide, dialkyl sulfoxide, diaryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfinate, aryl sulfinate, alkyl sulfonate, aryl sulfonate, sulfite, sulfate, thiosulfate, dialkyl selenide, diaryl selenide, alkyl aryl selenide, dialkyl selenoxide, diaryl selenoxide, alkyl aryl selenoxide, selenite, and seleninate; wherein Y and Z are each independently selected from the group consisting of hydroxide, alkoxide, aryloxide, siloxide, and halide;
(48) wherein n is 0 or 1;
(49) wherein m is 0 or 1; and
(50) wherein M, Y, Z, A.sup.1, A.sup.2, X.sup.1, X.sup.2, X.sup.3, X.sup.4, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and/or R.sup.17 are each independently optionally substituted.
(51) As used herein, a wavy bond indicates that the stereochemistry is unspecified, and encompasses all possible stereochemistries.
(52) In some instances, X.sup.1 and/or X.sup.2 are amino. For example, in doxaliplatin, both X.sup.1 and X.sup.2 are NH.sub.2. In some cases, X.sup.1 and X.sup.2 are joined together to form a bidentate ligand.
(53) In certain embodiments, X.sup.3 and/or X.sup.4 is a halide. In accordance with some embodiments, X.sup.3 and/or X.sup.4 is a carboxylate. For example, in doxaliplatin, both X.sup.3 and X.sup.4 are a carboxylate. X.sup.3 and X.sup.4 are joined together to form a bidentate ligand, in certain cases. For example, in doxaliplatin, X.sup.3 and X.sup.4 are joined together to form a bidentate ligand. According to some embodiments, X.sup.3 and X.sup.4 can be joined with one of X.sup.1 and X.sup.2 to form a tridentate ligand. Still further, in certain embodiments, both X.sup.3 and X.sup.4 can be joined with X.sup.1 and X.sup.2 to form a tetradentate ligand.
(54) In some cases, n and m are both 1. In other cases, n and m are both 0. For example, in doxaliplatin, n and m are both 0.
(55) In certain embodiments, M is selected from the group consisting of ammonium, tetra-alkyl ammonium, trialkylphenyl ammonium, lithium, sodium, potassium, magnesium, calcium, zinc, cobalt, copper, and iron.
(56) According to some embodiments, R.sup.2, R.sup.3, R.sup.4, and/or R.sup.13 are hydrogen.
(57) In accordance with certain embodiments, R.sup.6, R.sup.7, and/or R.sup.10 are hydroxy.
(58) In some cases, R.sup.9 and/or R.sup.12 are methylene.
(59) A.sup.1, A.sup.2, R.sup.5, and/or R.sup.8 are oxygen, in some instances.
(60) R.sup.1, in certain cases, is alkoxy.
(61) In some embodiments, R.sup.11 is —(C═O)—(CH.sub.2).sub.kR.sup.15.
(62) R.sup.15 is hydroxy, in some cases.
(63) In certain instances, k is 1.
(64) According to certain embodiments, R.sup.2, R.sup.3, R.sup.4, and/or R.sup.13 are hydrogen; A.sup.1, A.sup.2, R.sup.5, and/or R.sup.8 are oxygen; R.sup.1 is alkoxy; R.sup.11 is —(C═O)—(CH.sub.2).sub.kR.sup.15; R.sup.15 is hydroxy; and/or k is 1.
(65) In some embodiments, the compound of Formula (I) or Formula (II) is;
(66) ##STR00007##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.
(67) In certain embodiments, the compound of Formula (I) or Formula (II) is:
(68) ##STR00008##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.
(69) According to some embodiments, the compound of Formula (I) or Formula (II) is:
(70) ##STR00009##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof.
(71) In some embodiments, the compound is a compound of Formula (III):
(72) ##STR00010##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof;
(73) wherein A comprises at least 70% of the structure of an anthracycline drug;
(74) wherein X.sup.1 is selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, and diarylamino;
(75) wherein X.sup.2, X.sup.3, and X.sup.4 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, heteroarylene, water, halide, carboxylate, hydroxide, alkoxide, aryloxide, siloxide, dialkyl sulfide, diaryl sulfide, alkyl aryl sulfide, dialkyl sulfoxide, diaryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfinate, aryl sulfinate, alkyl sulfonate, aryl sulfonate, sulfite, sulfate, thiosulfate, dialkyl selenide, diaryl selenide, alkyl aryl selenide, dialkyl selenoxide, diaryl selenoxide, alkyl aryl selenoxide, selenite, and seleninate; and
(76) wherein A, X.sup.1, X.sup.2, X.sup.3, and/or X.sup.4 are each independently optionally substituted.
(77) In certain embodiments, embodiments relating to A, X.sup.1, X.sup.2, X.sup.3, and/or X.sup.4 described for Formula (I) apply to Formula (III).
(78) In accordance with some embodiments, the compound is a compound of Formula (IV):
(79) ##STR00011##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof;
(80) wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.6, and R.sup.7 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and—OM;
(81) wherein M is a cation;
(82) wherein R.sup.5 and R.sup.8 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM;
(83) wherein R.sup.9 and R.sup.12 are each independently selected from the group consisting of —CR(R′)—. carbonyl, imine, alkylimine, and arylimine;
(84) wherein R and R′ are independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, and diarylamino;
(85) wherein R.sup.10 and R.sup.11 are each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, —(C═O)—(CH.sub.2).sub.kR.sup.15, —(CHOR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(C═NR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(CHNHR.sup.16)—(CH.sub.2).sub.kR.sup.15, and —(CHNR.sup.16.sub.2)—(CH.sub.2).sub.kR.sup.15;
(86) wherein R.sup.15 is selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, and siloxy;
(87) wherein R.sup.16 is selected from the group consisting of hydrogen, alkyl, aryl, and silyl;
(88) wherein k is 0, 1, 2, or 3;
(89) wherein R.sup.13 and R.sup.14 are each independently selected from the group consisting of hydrogen, halogen, alkyl, and aryl;
(90) wherein A.sup.1 and A.sup.2 are each independently selected from the group consisting of oxygen, sulfur, and —NR.sup.17;
(91) wherein R.sup.17 is selected from the group consisting of hydrogen, alkyl, and aryl;
(92) wherein X.sup.1 is selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino;
(93) wherein X.sup.2 is selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, siloxy, alkyl, aryl, azido, and amide;
(94) wherein X.sup.3, X.sup.4, and X.sup.5 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, heteroarylene, water, halide, carboxylate, hydroxide, alkoxide, siloxide, dialkyl sulfide, diaryl sulfide, alkyl aryl sulfide, dialkyl sulfoxide, diaryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfinate, aryl sulfinate, alkyl sulfonate, aryl sulfonate, sulfite, sulfate, thiosulfate, dialkyl selenide, diaryl selenide, alkyl aryl selenide, dialkyl selenoxide, diaryl selenoxide, alkyl aryl selenoxide, selenite, and seleninate; and
(95) wherein M, A.sup.1, A.sup.2, X.sup.1, X.sup.2, X.sup.3, X.sup.4, X.sup.5, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and/or R.sup.17 are each independently optionally substituted.
(96) In certain embodiments, M is selected from the group consisting of ammonium, tetra-alkyl ammonium, trialkylphenyl ammonium, lithium, sodium, potassium, magnesium, calcium, zinc, cobalt, copper, and iron.
(97) In some cases, any combination of two of X.sup.3, X.sup.4, and X.sup.5 can be joined together to form a bidentate ligand.
(98) In certain instances, X.sup.3, X.sup.4, and X.sup.5 can be joined together to form a tridentate ligand.
(99) In certain cases, any of X.sup.3, X.sup.4, and X.sup.5 can be joined with X.sup.1 to form a bidentate, ligand.
(100) In some embodiments, any combination of two of X.sup.3, X.sup.4, and X.sup.5 can be joined with X.sup.1 to form a tridentate ligand.
(101) In certain embodiments, X.sup.3, X.sup.4, and X.sup.5 can be joined with X.sup.1 to form a tetradentate ligand.
(102) In certain embodiments, embodiments relating to M, A.sup.1, A.sup.2, X.sup.1, X.sup.2, X.sup.3, X.sup.4, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and/or R.sup.17 described for Formula (II) apply to Formula (IV).
(103) In accordance with some embodiments, the compound is a compound of Formula (V):
(104) ##STR00012##
or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof;
(105) wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.6, and R.sup.7 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM;
(106) wherein M is a cation;
(107) wherein R.sup.5 and R.sup.8 are each independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, oxygen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, imine, alkylimine, arylimine, and —OM;
(108) wherein R.sup.9 and R.sup.12 are each independently selected from the group consisting of —CR(R′)—, carbonyl, imine, alkylimine, and arylimine;
(109) wherein R and R′ are independently selected from the group consisting of hydrogen, alkyl, aryl, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, and diarylamino;
(110) wherein R.sup.10 and R.sup.11 are each independently selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, siloxy, amino, alkylamino, arylamino, dialkylamino, diarylamino, —(C═O)—(CH.sub.2).sub.kR.sup.15, —(CHOR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(C═NR.sup.16)—(CH.sub.2).sub.kR.sup.15, —(CHNHR.sup.16)—(CH.sub.2).sub.kR.sup.15, and —(CHNR.sup.16.sub.2)—(CH.sub.2).sub.kR.sup.15; wherein R.sup.15 is selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, and siloxy;
(111) wherein R.sup.16 is selected from the group consisting of hydrogen, alkyl, aryl, and silyl;
(112) wherein k is 0, 1, 2, or 3;
(113) wherein R.sup.13 and R.sup.14 are each independently selected from the group consisting of hydrogen, halogen, alkyl, and aryl;
(114) wherein A.sup.1 and A.sup.2 are each independently selected from the group consisting of oxygen, sulfur, and —NR.sup.17;
(115) wherein R.sup.17 is selected from the group consisting of hydrogen, alkyl, and aryl;
(116) wherein X.sup.1 is selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, and heteroarylene;
(117) wherein X.sup.2 is selected from the group consisting of hydrogen, halogen, hydroxy, alkoxy, aryloxy, siloxy, alkyl, aryl, azido, and amide;
(118) wherein X.sup.3, X.sup.4, and X.sup.5 are each independently selected from the group consisting of amino, alkylamino, arylamino, dialkylamino, diarylamino, heteroarylene, water, halide, carboxylate, hydroxide, alkoxide, aryloxide, siloxide, dialkyl sulfide, diaryl sulfide, alkyl aryl sulfide, dialkyl sulfoxide, diaryl sulfoxide, alkyl aryl sulfoxide, alkyl sulfinate, aryl sulfinate, alkyl sulfonate, aryl sulfonate, sulfite, sulfate, thiosulfate, dialkyl selenide, diaryl selenide, alkyl aryl selenide, dialkyl selenoxide, diaryl selenoxide, alkyl aryl selenoxide, selenite, and seleninate; and
(119) wherein M, A.sup.1, A.sup.2, X.sup.1, X.sup.2, X.sup.3, X.sup.4, X.sup.5, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and/or R.sup.17 are each independently optionally substituted.
(120) In some cases, any combination of two of X.sup.3, X.sup.4, and X.sup.5 can be joined together to form a bidentate ligand.
(121) In certain embodiments, M is selected from the group consisting of ammonium, tetra-alkyl ammonium, trialkylphenyl ammonium, lithium, sodium, potassium, magnesium, calcium, zinc, cobalt, copper, and iron.
(122) In certain instances, X.sup.3, X.sup.4, and X.sup.5 can be joined together to form a tridentate ligand.
(123) In certain cases, any of X.sup.3, X.sup.4, and X.sup.5 can be joined with X.sup.1 to form a bidentate, ligand.
(124) In some embodiments, any combination of two of X.sup.3, X.sup.4, and X.sup.5 can be joined with X.sup.1 to form a tridentate ligand.
(125) In certain embodiments, X.sup.3, X.sup.4, and X.sup.5 can be joined with X.sup.1 to form a tetradentate ligand.
(126) In certain embodiments, embodiments relating to M, A.sup.1, A.sup.2, X.sup.1, X.sup.2, X.sup.3, X.sup.4, R, R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5, R.sup.6, R.sup.7, R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14, R.sup.15, R.sup.16, and/or R.sup.17 described for Formula (II) apply to Formula (V).
(127) According to some embodiments, the compound is a compound of Formulas (I)-(V) or a pharmaceutically acceptable salt thereof.
(128) Certain aspects relate to compositions. As used herein, a composition comprises an active compound (e.g., a platinum-containing compound disclosed herein) and an excipient (e.g., a pharmaceutically acceptable excipient). 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.
(129) In certain embodiments, the composition comprises a compound of Formulas (I)-(V) or a pharmaceutically acceptable salt, solvate, hydrate, polymorph, co-crystal, tautomer, stereoisomer, isotopically labeled derivative, or prodrug thereof. In some cases, the composition comprises a compound of Formulas (I)-(V) or a pharmaceutically acceptable salt thereof. In some instances, the composition further comprises a pharmaceutically acceptable excipient.
(130) In some embodiments of Formulas (I)-(V), X.sup.1, X.sup.2, X.sup.3, X.sup.4, and/or X.sup.5 may dissociate from the platinum center under suitable in vivo conditions.
(131) Certain aspects relate to methods of treating a disease in a subject in need thereof.
(132) In some embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound and/or composition disclosed herein.
(133) According to some embodiments, administering can be accomplished by implanting, absorbing, ingesting, injecting, or inhaling the compound and/or composition.
(134) A “therapeutically effective amount” of a compound described herein is an amount sufficient to provide a therapeutic benefit in the treatment of a condition or to delay or minimize one or more symptoms associated with the condition. A therapeutically effective amount of a compound means an amount of therapeutic agent, alone or in combination with other therapies, which provides a therapeutic benefit in the treatment of the condition. The term “therapeutically effective amount” can encompass an amount that improves overall therapy, reduces or avoids symptoms, signs, or causes of the condition, and/or enhances the therapeutic efficacy of another therapeutic agent. In some embodiments, a therapeutically effective amount refers to an amount sufficient to treat a disease.
(135) The terms “treatment,” “treat,” and “treating” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a disease described herein. In some embodiments, treatment may be administered after one or more signs or symptoms of the disease have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease. For example, treatment may be administered to a susceptible subject prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of exposure to a pathogen). Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence.
(136) In certain embodiments, the disease is a proliferative disease. Examples of proliferative diseases include cancer, inflammatory diseases, or autoimmune diseases. Exemplary cancers include, but are not limited to, acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma), appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g., cholangiocarcinoma), bladder cancer, women's gynecological cancers, 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)), blood cancers, 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., “Waldenstrom'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 lymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g, cutaneous T-cell lymphoma (CTCL) (e.g, mycosis fungiodes, 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 (MM)), 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.ka. 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 neuroendoctrine 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, cervical cancer, breast cancer, testicular cancer, lung cancer, and vulvar cancer (e.g., Paget's disease of the vulva). For example, in some embodiments, the cancer is ovarian cancer and/or colorectal cancer.
(137) Examples of possible subjects include, but are not limited to, humans (i.e., a male or female human of any age group, e.g., a pediatric subject (e.g, infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult, or senior adult)) and/or other non-human animals, for example, mammals (e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys); commercially relevant mammals such as cats, and/or dogs. In certain embodiments, the animal is a mammal. The animal may be a male or female and at any stage of development. A non-human animal may be a transgenic animal.
(138) In accordance with some embodiments, compounds and/or compositions disclosed herein have numerous applications and benefits. For example, in some cases, the compounds and/or compositions disclosed herein are more potent than other drugs. In certain instances, compounds and/or compositions disclosed herein can be used to treat diseases which are resistant to other drugs. In certain embodiments, cells uptake a higher amount of compounds and/or compositions disclosed herein compared to other drugs.
(139) The following examples are intended to illustrate certain embodiments of the present invention, but do not exemplify the full scope of the invention.
Example 1
(140) Oxaliplatin is an FDA-approved platinum agent used for cancer treatment. Structurally, oxaliplatin features a non-labile chelating R,R-1,2-diaminocyclohexane (DACH) ligand and a labile oxalate group, which dissociates in vivo. Doxorubicin is a chemotherapeutic drug used for treating a broad spectrum of cancers. Structurally, the sugar moiety of doxorubicin (DOX), called daunosamine, provides a versatile platform for platination.
(141) The cis 1,2-amino alcohol unit of the daunosamine was stereoselectively converted to a chelating trans 1,2-diamine motif (
Synthesis
(142) To achieve the desired 4′-dehydroxyamination of DOX, the 3′-amino group and the 9- and 14-hydroxyl groups of DOX were protected. The 3′-amino group was protected with an N-trifluoroacetyl group. A cyclic orthoester was introduced to mask both 9- and 14-hydroxyl groups, essentially a 1,3-diol moiety.
(143) The synthesis started with the protection of the 3′-amino group by treating DOX hydrochloride with trifluoroacetic anhydride in pyridine (
(144) Cytotoxicity Studies
(145) The obtained platinum complex, doxaliplatin, displayed potent anticancer activity across a series of murine colorectal cancer organoids engineered to harbor mutations commonly found in human colon cancer. This organoid series was also engineered to express the fluorescent protein TdTomato, which can facilitate future in vivo studies, and comprises the following genotypes: 1. APC.sup.−/−; TdTomato.sup.+ (A-tdt), 2. APC.sup.−/−; P53.sup.−/−; TdTomato.sup.+ (AP-tdt), 3. APC.sup.−/−; KRAS.sup.G12D/WT; P53.sup.−/−; TdTomato.sup.+ (AKP-tdt), 4. APC.sup.−/−; KRAS.sup.G12D/WT; P53.sup.−/− (AKP), 5. APC.sup.−/−; KRAS.sup.G12D/WT; P53.sup.−/−; MSH2.sup.−/− (AKP-MSH2). Within the series of A-tdt, AP-tdt, and AKP-tdt, doxaliplatin exhibited three to ten-fold higher cellular toxicity than oxaliplatin did against these organoids (
(146) TABLE-US-00001 TABLE 1 IC.sub.50 values (μM) and the corresponding 95% confidence intervals (μM, in parentheses) of oxaliplatin (Ox), doxaliplatin (DoxPt), and doxorubicin (DOX) against various murine colorectal cancer organoids after 72-h exposure. A-tdt organoids AP-tdt organoids AKP-tdt organoids Ox 2.2 (1.9-2.4) 1.5 (1.2-2.0) 4.1 (3.4-5.0) DoxPt 0.46 (0.41-0.51) 0.41 (0.30-0.58) 0.43 (0.30-0.65) DOX 0.058 (0.051-0.065) 0.045 (0.035-0.059) 0.075 (0.059-0.097)
(147) The efficacy of doxaliplatin in the context of microsatellite unstable colon cancer was then investigated, using murine AKP-MSH2 colorectal cancer organoids as a model for microsatellite unstable colon cancers. Compared to AKP control organoids, AKP-MSH2 organoids were modestly resistant to oxaliplatin, but displayed increased sensitivity to doxaliplatin and doxorubicin (
(148) TABLE-US-00002 TABLE 2 IC.sub.50 values (μM) and the corresponding 95% confidence intervals (μM, in parentheses) of oxaliplatin (Ox), doxaliplatin (DoxPt), and doxorubicin (DOX) against AKP and AKP-MSH2 colorectal cancer organoids after 72-h exposure. AKP organoids AKP-MSH2 organoids Ox 1.4 (1.1-1.8) 2.2 (1.8-2.6) DoxPt 0.11 (0.091-0.13) 0.078 (0.058-0.10) DOX 0.0091 (0.0066-0.011) 0.0065 (0.0051-0.0078)
(149) The activity of doxaliplatin was studied against a panel of cell lines (
(150) In addition, an oxaliplatin-resistant ovarian cancer line, OVCAR8-OXR, was generated by exposing the parental OVCAR8 cells repeatedly with oxaliplatin. As shown in
(151) TABLE-US-00003 TABLE 3 IC.sub.50 values (μM) of oxaliplatin (Ox), doxaliplatin (DoxPt), doxorubicin (DOX), and a physical mixture of oxaliplatin and doxorubicin (DOX + Ox) against various cancer cell lines after 96-h exposure. MDA- HL60 HT-29 MB-231 PC3 U2OS A549 (leu- (colon) (breast) (prostate) (bone) (lung) kemia) Ox 0.47 0.92 1.0 0.58 0.40 0.56 DoxPt 0.33 0.11 0.092 0.052 0.10 0.16 DOX 0.038 0.075 0.041 0.013 0.023 0.043 DOX + Ox 0.046 — 0.048 0.012 0.020 0.055
(152) TABLE-US-00004 TABLE 4 IC.sub.50, IC.sub.90, and IC.sub.95 values (μM) of oxaliplatin (Ox), doxaliplatin (DoxPt), doxorubicin (DOX), and a physical mixture of oxaliplatin and doxorubicin (DOX + Ox) against the A2780 human ovarian cancer cell line after 72-h exposure. IC.sub.50 IC.sub.90 IC.sub.95 Ox 0.17 0.59 5.6 DoxPt 0.016 0.38 0.68 DOX 0.011 0.075 0.16 DOX + Ox 0.011 0.083 0.18
(153) TABLE-US-00005 TABLE 5 IC.sub.50, IC.sub.90, and IC.sub.95 values (μM) of oxaliplatin (Ox), doxaliplatin (DoxPt), doxorubicin (DOX), and a physical mixture of oxaliplatin and doxorubicin (DOX + Ox) against the doxorubicin-resistant A2780ADR human ovarian cancer cell line after 72-h exposure. IC.sub.50 IC.sub.90 IC.sub.95 Ox 0.43 2.4 29 DoxPt 0.072 0.49 0.67 DOX 0.054 0.62 1.6 DOX + Ox 0.053 0.51 2.4
(154) TABLE-US-00006 TABLE 6 IC.sub.50, IC.sub.90, and IC.sub.95 values (μM) of oxaliplatin (Ox), doxaliplatin (DoxPt), doxorubicin (DOX), and a physical mixture of oxaliplatin and doxorubicin (DOX + Ox) against the cisplatin-resistant A2780CIS human ovarian cancer cell line after 72-h exposure. IC.sub.50 IC.sub.90 IC.sub.95 Ox 0.43 6.6 13 DoxPt 0.28 0.38 0.41 DOX 0.050 0.39 0.54 DOX + Ox 0.049 0.36 0.49
(155) TABLE-US-00007 TABLE 7 IC.sub.50 values (μM) of oxaliplatin (Ox), doxaliplatin (DoxPt), doxorubicin (DOX), and a physical mixture of oxaliplatin and doxorubicin (DOX + Ox) against the OVCAR8 and OVCAR8-OXR human ovarian cancer cell line after 72-h exposure. IC.sub.50 (μM) OVCAR8 OVCAR8OXR Fold Resistance DOX 0.086 ± 0.003 0.25 ± 0.01 2.9 Ox 0.58 ± 0.06 8.2 ± 1.3 14 DOX + Ox 0.10 ± 0.01 0.27 ± 0.01 2.7 DoxPt 0.14 ± 0.01 0.14 ± 0.01 1.0
(156) TABLE-US-00008 TABLE 8 IC.sub.80 values (μM) of oxaliplatin (Ox), doxaliplatin (DoxPt), doxorubicin (DOX), and a physical mixture of oxaliplatin and doxorubicin (DOX + Ox) against the OVCAR8 and OVCAR8-OXR human ovarian cancer cell line after 72-h exposure. IC.sub.80 (μM) OVCAR8 OVCAR8OXR Fold Resistance DOX 0.41 ± 0.02 0.74 ± 0.05 1.8 Ox 12 ± 3 40 ± 2 3.3 DOX + Ox 0.41 ± 0.02 0.78 ± 0.08 1.9 DoxPt 0.37 ± 0.02 0.23 ± 0.01 0.6
Platinum Uptake
(157) The whole-cell platinum uptake of oxaliplatin and doxaliplatin was investigated under different conditions using inductively coupled plasma mass spectrometry (ICP-MS). As shown in
(158) TABLE-US-00009 TABLE 9 Platinum drug uptake (nmol/mg protein) by A2780 human ovarian cancer cell line and its doxorubicin-resistant variant A2780ADR at drug concentrations during the treatment. A2780 A2780 A2780ADR A2780ADR [Drug] (μM) treated with Ox treated with DOX treated with Ox treated with DOX 0.00 0.0020 ± 0.0002 0.00038 ± 0.00045 0.20 0.0029 ± 0.0006 0.023 ± 0.002 0.0011 ± 0.0004 0.0087 ± 0.0008 1.0 0.0093 ± 0.0004 0.12 ± 0.01 0.0039 ± 0.0001 0.051 ± 0.001 5.0 0.050 ± 0.004 0.63 ± 0.03 0.023 ± 0.001 0.36 ± 0.07
(159) It was further explored whether high drug accumulation of the covalent conjugate, doxaliplatin, could be attained by using a physical mixture of doxorubicin and oxaliplatin. In this separate experiment, the platinum content in A2780 cells and A2780ADR cells treated with oxaliplatin alone or a 1:1 combination of oxaliplatin and doxorubicin were compared (Table 10). As shown in
(160) TABLE-US-00010 TABLE 10 Comparison of platinum drug uptake (nmol/mg protein) of doxaliplatin, oxaliplatin alone, and a physical mixture of oxaliplatin with doxorubicin. Drug A2780 A2780ADR untreated 0.00057 ± 0.00013 0.00027 ± 0.00009 Ox 0.011 ± 0.000 0.0065 ± 0.0004 Ox + DOX 0.014 ± 0.005 0.0070 ± 0.0000 DoxPt 0.089 ± 0.031 0.061 ± 0.003
CONCLUSION
(161) In summary, a synthetic route that facilitated the platination of a doxorubicin derivative through a non-cleavable bidentate binding mode was demonstrated. Exploiting this synthetic protocol, a novel platinum agent, doxaliplatin, which displayed a significantly enhanced anticancer profile compared to oxaliplatin and doxorubicin, singly or in combination, was obtained. The improvement was particularly substantial when treating doxorubicin- or oxaliplatin-resistant cell lines. It is unequivocal that the synthetic schemes devised en route to doxaliplatin may facilitate conjugation of other anthracyclines to platinum agents used in the clinic.
(162) Experimental Section
(163) Materials and Methods.
(164) Reagents were purchased from commercial sources and used as received. Doxorubicin hydrochloride was purchased from A Chemtek Inc. and its purity was examined by .sup.1H NMR spectroscopy and analytical HPLC. Anhydrous solvents were saturated with argon and purified by passage through two columns of activated alumina. Air-sensitive reactions and compounds were handled with standard Schlenk techniques or in an MBraun dry box. Column chromatography was performed on silica gel (230-400 mesh, 60 Å).
(165) The title platinum-containing compound was purified by a reverse phase Biotage® SNAP Ultra C18 column (12 g) using H.sub.2O/MeOH as the mobile phase prior to HPLC purification. Compound 9 (DOX-NH.sub.2—NH.sub.2) and doxaliplatin were purified using Agilent 1260 Series HPLC systems fitted with multi-wavelength detectors using a C18 reverse stationary phase (Zorbax-SB C18 column: 7 μm, 21.2×250 mm). The purity of compound 9 and doxaliplatin was examined using an Agilent 1200 Series HPLC system fitted with multi-wavelength detectors using a C18 reverse stationary phase (Zorbax-SB C18 column: 5 μm, 4.6×250 mm). The mobile phase was composed of two solvents. For 9, the mobile phase was composed of A: H.sub.2O+0.1% (v/v) CF.sub.3CO.sub.2H; B: CH.sub.3CN+0.1% (v/v) CF.sub.3CO.sub.2H. For doxaliplatin, the mobile phase was composed of A: H.sub.2O; B: CH.sub.3OH.
(166) NMR spectra were acquired on a 400 MHz Bruker AVANCE-400 spectrometer or a Varian Inova-500 NMR spectrometer. .sup.1H NMR and .sup.13C NMR chemical shifts are reported in ppm relative to that of SiMe.sub.4 (δ=0.00) and were referenced internally to residual solvent peaks..sup.31 195Pt NMR chemical shifts were reported in ppm relative to that of K.sub.2PtCl.sub.6 (δ=0.00). Low-resolution electrospray mass spectra were acquired on an Agilent 1100 Series LC/MSD Trap spectrometer. High-resolution mass spectra were acquired on an Agilent 6510 Series Quadrupole Time-Of-Flights spectrometer at the MIT Center for Environmental Health Sciences. MS/MS mass spectra were acquired on an Agilent 6410 Triple Quadrupole LC/MS at a fragmentor voltage of 202.0V and a collisionally induced dissociation of 40.0 eV at the MIT Center for Environmental Health Sciences.
(167) ICP-MS was performed on an Agilent 7900 ICP-MS in helium mode. The ICP-MS was equipped with an integrated auto-sampler at the MIT CEHS core facility.
(168) Colorectal Cancer Organoid Generation, and Testing of Oxaliplatin, Doxorubicin, and Doxaliplatin
(169) Colonic crypts were isolated and colon organoids were generated from the following mouse genotypes on a C57Bl/6 background: 1. Rosa-LSL-TdTomato (hereafter LSL-tdt), 2. P53.sup.f1/f1; LSL-tdt, and 3. LSL-KRAS.sup.G12D/WT; P53.sup.fl/fl; LSL-tdt mice. Subsequently, the pSECC-APC plasmid (carrying Cre, Cas9, and sgAPC) was transfected into each of the above three genotypes of colon organoids. This transfection strategy generated, in a single step, the following colorectal cancer organoids, including 1. APC.sup.−/−; TdTomato.sup.+ (A-tdt), 2. APC.sup.−/−; P53.sup.−/−; TdTomato.sup.+ (AP-tdt), and 3. APC.sup.−/−; KRAS.sup.G12D/WT; P53.sup.−/−; TdTomato.sup.+ (AKP-tdt). Organoids of the desired genotype were selected for by Wnt/Rspo withdrawal for APC.sup.−/−, Nutlin-3 addition for P53.sup.−/−, and EGF withdrawal for KRAS.sup.G12D.
(170) To generate 4. APC.sup.−/−; KRAS.sup.G12D/WT; P53.sup.−/−; (AKP) and 5. APC.sup.−/−; KRAS.sup.G12D/WT; P53.sup.−/−; MSH2.sup.−/− (AKP-MSH2) colon cancer organoids, colonic crypts were isolated and colon organoids were generated from the following mouse genotypes on a C57Bl/6 background: 4. LSL-KRAS.sup.G12D/WT; P53.sup.fl/fl mice and 5. LSL-KRAS.sup.G12D; P53.sup.fl/fl; MSH2.sup.fl/fl mice. Subsequently, the pSECC-APC plasmid (carrying Cre, Cas9, and sgAPC) was transfected into each of the above two genotypes of colon organoids. Following pSECC-APC transfection, Wnt/Rspo withdrawal was performed and surviving (APC.sup.−/−) single organoids were cloned and PCR genotyped for Cre mediated deletion of p53 and/or MSH2, and for Cre mediated activation of oncogenic KRAS.sup.G12D.
(171) To determine IC.sub.50 values for oxaliplatin, doxorubicin, and doxaliplatin in A-tdt, AP-tdt, and AKP-tdt organoids, organoids were trypsinized and seeded in 10 μL drops of 67% Corning Matrigel (growth factor reduced) with 33% culture media described below. Twenty four hours after seeding the organoids, a media change was performed and dose responses of oxaliplatin, doxorubicin and doxaliplatin were added. Forty eight hours after treatment, cell growth was evaluated by resazurin assay. Organoids were cultured in Advanced DMEM/F12 supplemented with 2 mM GlutaMAX, 1% Penicillin-Streptomycin, 2% B27, and Y-27632 (10 μM).
(172) Testing Oxaliplatin, Doxorubicin, and Doxaliplatin in Cell Lines
(173) Cells were cultured in 10 mL of Advanced DMEM/F12 (ADMEM/F12), supplemented with 5% FBS, 2 mM GlutaMAX, and 1% Penicillin-Streptomycin in 10 cm tissue culture plates. Cells were then plated into 96-well plates with about 2.5×10.sup.3 cells per well. Cells were plated with 200 μL per well of ADMEM/F12 supplemented with 5% FBS, 2 mM GlutaMAX, and 1% Penicillin-Streptomycin. Cells were treated with dose responses of doxorubicin, oxaliplatin, or doxaliplatin upon plating. The cells were then cultured for 72 hours following drug treatment, and cell growth was measured by resazurin assay.
(174) Determining Platinum Uptake Using ICP-MS
(175) The cells in a six-well plate were treated with oxaliplatin, doxaliplatin, or a mixture of doxorubicin and oxaliplatin at indicated concentration in 2 mL of Advanced DMEM/F12 (ADMEM/F12), supplemented with 5% FBS, 2 mM GlutaMAX, and 1% Penicillin-Streptomycin. After three hours, the media were removed. The cells were washed with xl PBS buffer containing EDTA (100 μM, 4 mL×2). To each well, MilliQ water (1.0 mL) was added. After 15 min, the cells were suspended. A portion of the resulting cell lysate (900 μL) was transferred to an Eppendorf tube. The samples were frozen in liquid nitrogen and thawed at room temperature. This procedure was repeated for three times. The insoluble lysate was pelleted (10000 rpm for 10 min). The soluble protein concentration was determined using Pierce® 660 nm Protein assay in a 96-well plate as described below. Pierce 660 nm Protein solution (200 μL) was added to each well. An aliquot of the lysate (14.2 μL) was added to each well. The plate was shaken and read on a plate reader. The remaining samples were frozen at liquid nitrogen temperature and lyophilized. The dried material was digested with Aristar® ultrapure concentrated nitric acid (70%) at 70° C. for one hour. An aliquot of this solution (142 μL) was mixed with molecular biology grade water (2000 μL) and a terbium internal standard (40 μL, 100 ppb). The platinum content was then determined using an Agilent 7900 ICP-MS in helium mode. The concentration of platinum in the cells was normalized to measured protein concentration described above.
Synthetic Procedures
A-Trifluoroacetyl Doxorubicin (2)
(176) ##STR00013##
(177) In a Schlenk flask containing doxorubicin hydrochloride salt (1.16 g, 2.0 mmol), anhydrous pyridine (60 mL) was added at −30° C. The reaction was stirred at this temperature for 15 min. A mixture of trifluoroacetic anhydride (3.78 g, 2.54 mL, 18.0 mmol) in Et.sub.2O (12 mL) was added to the flask dropwise at the same temperature. The reaction was gradually warmed to 0° C. over a period of 3 h, during which doxorubicin dissolved to give a slightly brown solution. The reaction was diluted with water (100 mL) and stirred for 20 min to hydrolyze excess trifluoroacetic anhydride. The reaction mixture was extracted with ethyl acetate (100 mL×3). The combined organic phase was washed with water (100 mL×3) and dried over MgSO.sub.4. The solvent was removed under vacuum. The crude product was suspended in a mixture of butanone and hexanes (80 mL+320 mL, respectively) and stored at −40° C. for 3 h. The dark red solid was collected by suction filtration, washed with hexanes, and dried under vacuum (1.20 g, 94% yield). .sup.1H and .sup.19F NMR spectra indicated that, presumably due to the slow amide bond rotation, two conformers were present in DMSO-d.sub.6 with a ratio of 4:1. .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 13.98 (s, 1.0H), 13.20 (s, 1.2H), 9.06-9.08 (m, 1.4H), 8.62-8.50 (m, 0.3H), 7.93-7.75 (m, 3.0H), 7.67-7.55 (m, 1.3H), 7.41-7.38 (m, 0.3H), 5.43 (s, 1.0H), 5.24-5.20 (s, 1.5H), 4.99 (d, J=5.9 Hz, 1.2H), 4.91 (dd, J=5.3, 2.8 Hz, 0.9H), 4.85 (t, J=5.9 Hz, 0.9H), 4.58 (d, J=5.9 Hz, 2.0H), 4.22 (q, J=6.3 Hz, 1.0H), 4.09-3.77 (m, 5.0H), 3.52 (d, J=3.9 Hz, 0.8H), 2.91 (dd, J=45.7, 18.2 Hz, 2.0H), 2.26-1.95 (m, 3.5H), 1.47 (dd, J=12.2, 4.1 Hz, 1.0H), 1.18-1.07 (m, 4.0H). .sup.19F NMR (396 MHz, DMSO-d.sub.6) −73.94 (s, 3F), −73.99 (s, 0.84F). ESI-MS(−) m/z calcd for [M−H].sup.− 638.1, found 638.0.
N-Trifluoroacetyl doxorubicin 9,14-cyclic methyl orthoester (3)
(178) ##STR00014##
(179) To a solution of 2 (1.06 g, 1.66 mmol) and methyl orthoformate (80 mL) in THF (280 mL) was added (1S)-(+)-10-camphorsulfonic acid (102 mg, 0.44 mmol) in one portion. The reaction was stirred at room temperature for 3 h. The reaction was then quenched with 5 wt % NaHCO.sub.3 aq. solution (400 mL). The mixture was extracted with ethyl acetate (100 mL×3). The combined organic phase was washed with 5 wt % NaHCO.sub.3 aq. solution (200 mL) and water (200 mL). The organic phase was dried over MgSO.sub.4. The solvent was removed under vacuum. The crude product was purified by column chromatography (silica gel, ethyl acetate:hexanes=3:1 to 6:1) to afford a dark orange solid (362 mg, 32% yield). ESI-MS(−) m/z calcd for [M−H].sup.− 680.2, found 680.1. .sup.19F NMR spectrum of 3 in acetone-d.sub.6 displayed four major signals with a ratio of 70:18:7:5. These signals appeared to correspond to two diastereomers of the cyclic methyl orthoester, each of which has two conformers due to the slow amide bond rotation.
N-Trifluoroacetyl (4′S)-triflate doxorubicin 9,14-cyclic methyl orthoester (4)
(180) ##STR00015##
(181) To a Schlenk flask containing 3 (301 mg, 0.44 mmol) in CH.sub.2Cl.sub.2 (10 mL), pyridine (280 mg, 285 μL, 3.54 mmol) was added dropwise at −40° C. Triflic anhydride (499 mg, 297 μL, 1.77 mmol) in CH.sub.2Cl.sub.2 (5 mL) was then added dropwise to the flask at the same temperature. The reaction was stirred for 2 h, during which the reaction was gradually warmed to 0° C. The completion of the reaction was confirmed by thin layer chromatography (ethyl acetate:hexanes=3:1). The reaction was diluted with ice water (40 mL). The mixture was extracted with cold Et.sub.2O (40 mL×3). The combined organic phase was washed with cold NaOAc aq. solution (10 wt %, 80 mL) and then cold water (80 mL). The organic phase was dried over MgSO.sub.4. The solvent was removed under vacuum. The crude product (346 mg) was used in the next step without further purification.
N-Trifluoroacetyl (4′R)-azido doxorubicin 9,14-cyclic methyl orthoester (5)
(182) ##STR00016##
(183) In the glovebox, 4 (crude, 346 mg, approximately 0.43 mmol) was dissolved in anhydrous THF (15) mL). To this solution, nBu.sub.4NN.sub.3 (121 mg, 0.43 mmol) was added. A dark red solution formed immediately. The reaction was stirred at room temperature for 12 h. The solvent was removed under vacuum. The crude product was purified by column chromatography (ethyl acetate:hexanes=2:1). An orange solid was obtained (199 mg, 64% yield based on 3). ESI-MS(−) m/z calcd for [M−H].sup.− 705.2, found 705.1.
(4′R)-Azido doxorubicin 9,14-cyclic methyl orthoester (6)
(184) ##STR00017##
(185) To a stirred solution of 5 (199 mg, 0.28 mmol) in CHCl.sub.3 (140 mL) was added NaOH in sat. brine (0.5 M, 14.0 mL). The reaction was stirred vigorously (1200 rpm) at room temperature for 24 h. The completion of the reaction was confirmed by ESI-MS. The organic phase was isolated. The aqueous phase was extracted with CH.sub.2Cl.sub.2 (50 mL×2). The combined organic phase was washed with brine (50 mL×2) and water (50 mL×2). The organic phase was dried over MgSO.sub.4. The solvent was removed under vacuum. The crude product was used in the next step without further purification (80 mg, 47% yield). ESI-MS(−) m/z calcd for [M−H].sup.− 609.2, found 609.2. ESI-MS(+) m/z calcd for [M+H].sup.+611.2, found 611.2.
(4′R)-Dehydroxyamino doxorubicin (DOX-NH.SUB.2.—NH.SUB.2., 9)
(186) ##STR00018##
(187) To the crude product 6 (80 mg, 0.13 mmol) in THF (52 mL) was added triphenylphosphine (102 mg, 0.39 mmol) in small portions. The reaction was stirred at room temperature for 48 h. The completion of the reaction was confirmed by ESI-MS. ESI-MS(+) m/z calcd for iminophosphorane (7) [M+H].sup.+ 845.3, found 845.5.
(188) The above reaction was mixed with water (5.2 mL) and stirred at room temperature. The progress of the hydrolysis of iminophosphorane (7) was monitored by ESI-MS, which was complete after 24 h. ESI-MS(+) m/z calcd for (4′R)-dehydroxyamino doxorubicin 9,14-cyclic methyl orthoester (8) [M+H].sup.+ 585.2, found 585.1; and [M+Ph.sub.3PO+H].sup.+863.3, found 863.3.
(189) To the same reaction mixture, an aliquot of aq. HCl (0.25 M, 1.04 mL, 0.26 mmol) was added. The progress of the reaction was monitored by ESI-MS. The reaction was complete after 24 h. The solvents were removed under vacuum. The reaction mixture was dissolved in MeCN/water (1:1, v/v, 20 mL). The mixture was first suction filtered. The filtrate was then passed through a 0.2 μm PTFE syringe filter. The crude product was purified by preparative HPLC. Preparative HPLC used the (A) water (0.1% v/v CF.sub.3CO.sub.2H)/(B) CH.sub.3CN (0.1% v/v CF.sub.3CO.sub.2H) solvent system, according to the following protocol: constant flow rate 15.0 mL.Math.min.sup.−1; 0.0-3.0 min, linear gradient 20-27% B; 3.0-13.0 min, linear gradient 27-37% B; 13.0-15.0 min, linear gradient 37-100% B; 15.0-18.0 min, 100% B; 18.0-20.0 min, linear gradient 100-20% B; 20.0-21.0 min, 20% B. Fractions containing the desired product (T.sub.R=11.8 min) were combined and lyophilized to give an orange solid as a bis(trifluoroacetic acid) salt (48 mg, 48% yield based on 6). The purity of 9 was examined by analytical HPLC (T.sub.R=17.2 min). Analytical HPLC used the (A) water (0.1% v/v CF.sub.3CO.sub.2H)/(B) CH.sub.3CN (0.1% v/v CF.sub.3CO.sub.2H) solvent system, according to the following protocol: constant flow rate 1.0 mL.Math.min.sup.−1; 0.0-5.0 min, 10% B; 5.0-30.0 min, linear gradient 10-100% B; 30.0-33.0 min, 100% B; 33.0-36.0 min, linear gradient 100-10% B; 36.0-40.0 min, 10% B. UV-Vis (50 mM PIPES, 100 mM KCl, pH 7.0) ε.sub.490=18,300 M.sup.−1.Math.cm.sup.−1. The .sup.1H NMR spectrum of 9 in DMSO-d.sub.6 displayed multiple sets of signals, a feature indicating the presence of multiple species. In contrast, the .sup.1H NMR spectrum of 9 in CD.sub.3CN/D.sub.2O (3:1, v/v) only showed one set of signals. This observation suggested that compound 9 underwent slow conformational interconversions on the NMR timescale in DMSO-d.sub.6. .sup.1H NMR (400 MHz, CD.sub.3CN/D.sub.2O 3:1, v/v) δ 7.76-7.64 (m, 2H), 7.41 (d, J=9.0 Hz, 1H), 5.45 (d, J=2.8 Hz, 1H), 4.94 (d, J=2.7 Hz, 1H), 4.69 (d, J=20 Hz, 2H, AB system), 4.16 (dq, J=10.0, 6.3 Hz, 1H), 3.91 (s, 3H), 3.67 (ddd, J=12.0, 10.5, 4.6 Hz, 1H), 3.15 (t, J=10.0 Hz, 1H), 3.00 (d, J=19.2 Hz, 1H, part of an AB system), 2.74 (d, J=18.7 Hz, 1H, part of an AB system), 2.32-2.04 (m, 3H), 1.94 (1H, overlap with CD.sub.2HCN), 1.36 (d, J=6.3 Hz, 3H). .sup.13C{.sup.1H} NMR (101 MHz, CD.sub.3CN/D.sub.2O 3:1, v/v) δ 214.8, 187.7, 187.6, 161.9, 156.7, 155.4, 137.3, 135.6, 134.7 (2), 134.7 (0), 120.8, 120.4, 120.3, 112.2, 112.0, 99.3, 76.7, 70.4, 66.3, 65.5, 57.4, 55.6, 47.6, 36.8, 34.1, 33.2, 17.9. ESI-MS(+) m/z calcd for DOX-NH.sub.2—NH.sub.2 (9) [M+H].sup.+ 543.2, found 543.4. ESI-MS(−) m/z calcd [M−H].sup.− 541.2, found 541.2. ESI-HRMS(+) m/z calcd for C.sub.27H.sub.31N.sub.2O.sub.10.sup.+[M+H].sup.+ 543.1973, found 543.1947.
Doxaliplatin (12)
(190) ##STR00019##
(191) To a stirred solution of K.sub.2PtCl.sub.4 (41.5 mg, 0.100 mmol) in water (30 mL), a solution of 9 bis(trifluoroacetic acid) salt (77.0 mg, 0.100 mmol) in water (9 mL) was added in one portion. Red precipitate formed immediately. The reaction was stirred at room temperature for 24 h. To the same solution was added AgNO.sub.3 aq. solution (0.10 M, 4.0 mL). The reaction was stirred at room temperature for 24 h. To this mixture was then added K.sub.2C.sub.2O.sub.4 aq. solution (0.20 M, 0.50 mL). The reaction was stirred at room temperature for 24 h. The reaction mixture was lyophilized together with a small amount of C18-reversed phase silica gel. The crude product was first purified using a reverse phase Biotage® SNAP Ultra C18 column (12 g). The purification used the (A) water/(B) MeOH solvent system, according to the following protocol: constant flow rate 12.0 mL.Math.min.sup.−1; 0.0-3.0 column volume (CV), 0% B; 3.0-10.0 CV, linear gradient 0-80% B; 10.0-20.0 CV, 80% B. Fractions from 13-15 CV were collected and filtered through a 0.2 μm PTFE syringe filter. The combined fractions were further purified by preparative HPLC. Preparative HPLC used the (A) water/(B) MeOH solvent system, according to the following protocol: constant flow rate 15.0 mL.Math.min.sup.−1; 0.0-3.0 min, linear gradient 25-62% B; 3.0-13.0 min, linear gradient 62-72% B; 13.0-15.0 min, linear gradient 62-100% B; 15.0-17.0 min, 25% B; 17.0-18.0 min, 25% B. Fractions containing the desired product (T.sub.R=12.8 min) were combined and lyophilized to give a bright orange solid (24.0 mg, 29% yield based on 9). The purity of 12 was examined by analytical HPLC (T.sub.R=25.7 min). Analytical HPLC used the (A) water/(B) MeOH solvent system, according to the following protocol: constant flow rate 1.0 mL.Math.min.sup.−1; 0.0-5.0 min, 10% B; 5.0-30.0 min, linear gradient 10-100% B; 30.0-33.0 min, 100% B; 33.0-36.0 min, linear gradient 100-10% B; 36.0-40.0 min, 10% B. UV-Vis (water) ε.sub.478=16,500 M.sup.−1.Math.cm.sup.−1. The .sup.1H NMR spectrum of 12 in DMF-d.sub.7 showed a set of signals with significant line broadening, which indicated that the observed species underwent a slow exchange reaction. The low solubility of 12 in DMF, however, impeded the characterization of the compound. The stability and the relatively high solubility.sup.32 of 12 in DMSO-d.sub.6 allowed further characterization of the compound. In contrast to the .sup.1H NMR spectrum in DMF-d.sub.7, the .sup.1H NMR spectrum of 12 in DMSO-d.sub.6 revealed two sets of signals, presumably due to the presence of two conformers (
(192) While several embodiments of the present invention have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and/or structures for performing the functions and/or obtaining the results and/or one or more of the advantages described herein, and each of such variations and/or modifications is deemed to be within the scope of the present invention. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the teachings of the present invention is/are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto, the invention may be practiced otherwise than as specifically described and claimed. The present invention is directed to each individual feature, system, article, material, and/or method described herein. In addition, any combination of two or more such features, systems, articles, materials, and/or methods, if such features, systems, articles, materials, and/or methods are not mutually inconsistent, is included within the scope of the present invention.
(193) The indefinite articles “a” and “an,” as used herein in the specification and in the claims, unless clearly indicated to the contrary, should be understood to mean “at least one.”
(194) The phrase “and/or,” as used herein in the specification and in the claims, should be understood to mean “either or both” of the elements so conjoined, i.e., elements that are conjunctively present in some cases and disjunctively present in other cases. Other elements may optionally be present other than the elements specifically identified by the “and/or” clause, whether related or unrelated to those elements specifically identified unless clearly indicated to the contrary. Thus, as a non-limiting example, a reference to “A and/or B,” when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A without B (optionally including elements other than B); in another embodiment, to B without A (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.
(195) As used herein in the specification and in the claims, “or” should be understood to have the same meaning as “and/or” as defined above. For example, when separating items in a list, “or” or “and/or” shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as “only one of” or “exactly one of,” or, when used in the claims, “consisting of,” will refer to the inclusion of exactly one element of a number or list of elements. In general, the term “or” as used herein shall only be interpreted as indicating exclusive alternatives (i.e. “one or the other but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “only one of,” or “exactly one of.” “Consisting essentially of,” when used in the claims, shall have its ordinary meaning as used in the field of patent law.
(196) As used herein in the specification and in the claims, the phrase “at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase “at least one” refers, whether related or unrelated to those elements specifically identified. Thus, as a non-limiting example, “at least one of A and B” (or, equivalently, “at least one of A or B,” or, equivalently “at least one of A and/or B”) can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.
(197) As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al, describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid or by using other methods known in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N.sup.+(C.sub.1-4 alkyl).sub.4.sup.− salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.
(198) Optionally substituted refers to a group which may be substituted or unsubstituted. In general, the term “substituted” means that at least one hydrogen present on a group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a “substituted” group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term “substituted” is contemplated to include substitution with all permissible substituents of organic compounds, and includes any of the substituents described herein that results in the formation of a stable compound. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any suitable substituent as described herein which satisfy the valencies of the heteroatoms and results in the formation of a stable moiety.
(199) Exemplary carbon atom substituents include, but are not limited to, halogen, —CN, —NO.sub.2, —N.sub.3, —SO.sub.2H, —SO.sub.3H, —OH, —OR.sup.aa, —ON(R.sup.bb).sub.2, —N(R.sup.bb).sub.2, —N(R.sup.bb).sub.3.sup.+X.sup.−, —N(OR.sup.cc)R.sup.bb, —SH, —SR.sup.aa, —SSR.sup.cc, —C(═O)R.sup.aa, —CO.sub.2H, —CHO, —C(OR.sup.cc).sub.2, —CO.sub.2R.sup.aa, —OC(═O)R.sup.aa, —OCO.sub.2R.sup.aa, —C(═O)N(R.sup.bb).sub.2, —OC(═O)N(R.sup.bb).sub.2, —NR.sup.bbC(═O)R.sup.aa, —NR.sup.bbCO.sub.2R.sup.aa, —NR.sup.bbC(═O)N(R.sup.bb).sub.2, —C(═NR.sup.bb)R.sup.aa, —C(═NR.sup.bb)OR.sup.aa, —OC(═NR.sup.bb)R.sup.aa, —OC(═NR.sup.bb)OR.sup.aa, —C(═NR.sup.bb)N(R.sup.bb).sub.2, —OC(═NR.sup.bb)N(R.sup.bb).sub.2, —NR.sup.bbC(═NR.sup.bb)N(R.sup.bb).sub.2, —C(═O)NR.sup.bbSO.sub.2R.sup.aa, —NR.sup.bbSO.sub.2R.sup.aa, —SO.sub.2N(R.sup.bb).sub.2, —SO.sub.2R.sup.aa, —SO.sub.2OR.sup.aa, —OSO.sub.2R.sup.aa, —S(═O)R.sup.aa, —OS(═O)R.sup.aa, —Si(R.sup.aa).sub.3, —OSi(R.sup.aa).sub.3—C(═S)N(R.sup.bb).sub.2, —C(═O)SR.sup.aa, —C(═S)SR.sup.aa, —SC(═S)SR.sup.aa, —SC(═O)SR.sup.aa, —OC(═O)SR.sup.aa, —SC(═O)OR.sup.aa, —SC(═O)R.sup.aa, —P(═O).sub.2R.sup.aa, —OP(═O).sub.2R.sup.aa, —P(═O)(R.sup.aa).sub.2, —OP(═O)(R.sup.aa).sub.2, —OP(═O)(OR.sup.cc).sub.2, —P(═O).sub.2N(R.sup.bb).sub.2, —OP(═O).sub.2N(R.sup.bb).sub.2, —P(═O)(NR.sup.bb).sub.2, —OP(═O)(NR.sup.bb).sub.2, —NR.sup.bbP(═O)(OR.sup.cc).sub.2, —NR.sup.bbP(═O)(NR.sup.bb).sub.2, —P(R.sup.cc).sub.2, —P(R.sup.cc).sub.3, —OP(R.sup.cc).sub.2, —OP(R.sup.cc).sub.3, —B(R.sup.aa).sub.2, —B(OR.sup.cc).sub.2, —BR.sup.aa(OR.sup.cc), C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, heteroC.sub.1-10 alkyl, heteroC.sub.2-10 alkenyl, heteroC.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
(200) or two geminal hydrogens on a carbon atom are replaced with the group ═O, ═S, ═NN(R.sup.bb).sub.2, ═NNR.sup.bbC(═O)R.sup.aa, ═NNR.sup.bbC(═O)OR.sup.aa, ═NNR.sup.bbS(═O).sub.2R.sup.aa, ═NR.sup.bb, or ═NOR.sup.cc;
(201) each instance of R.sup.aa is, independently, selected from C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, heteroC.sub.1-10 alkyl, heteroC.sub.2-10alkenyl, heteroC.sub.2-10alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14aryl, and 5-14 membered heteroaryl, or two R.sup.aa groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
(202) each instance of R.sup.bb is, independently, selected from hydrogen, —OH, —OR.sup.aa, —N(R.sup.cc).sub.2, —CN, —C(═O)R.sup.aa, —C(═O)N(R.sup.cc).sub.2, —CO.sub.2R.sup.aa, —SO.sub.2R.sup.aa, —C(═NR.sup.cc)OR.sup.aa, —C(═NR.sup.cc)N(R.sup.cc).sub.2, —SO.sub.2N(R.sup.cc).sub.2, —SO.sub.2R.sup.cc, —SO.sub.2OR.sup.cc, —SOR.sup.aa, —C(═S)N(R.sup.cc).sub.2, —C(═O)SR.sup.cc, —C(═S)SR.sup.cc, —P(═O).sub.2R.sup.aa, —P(═O)(R.sup.aa).sub.2, —P(═O).sub.2N(R.sup.cc).sub.2, —P(═O)(NR.sup.cc).sub.2, C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, heteroC.sub.1-10 alkyl, heteroC.sub.2-10alkenyl, heteroC.sub.2-10alkynyl, C3-10 carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two R.sup.bb groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
(203) each instance of R.sup.cc is, independently, selected from hydrogen, C.sub.1-10 alkyl, C.sub.1-10 perhaloalkyl, C.sub.2-10 alkenyl, C.sub.2-10 alkynyl, heteroC.sub.1-10 alkyl, heteroC.sub.2-10 alkenyl, heteroC.sub.2-10 alkynyl, C.sub.3-10 carbocyclyl, 3-14 membered heterocyclyl, C.sub.6-14 aryl, and 5-14 membered heteroaryl, or two R.sup.cc groups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.dd groups;
(204) each instance of R.sup.dd is, independently, selected from halogen, —CN, —NO.sub.2, —N.sub.3, —SO.sub.2H, —SO.sub.3H, —OH, —OR.sup.ee, —ON(R.sup.ff).sub.2, —N(R.sup.ff).sub.2, —N(R.sup.ff).sub.3.sup.+X.sup.−, —N(OR.sup.ee)R.sup.ff, —SH, —SR.sup.ee, —SSR.sup.ee, —C(═O)R.sup.ee, —CO.sub.2H, —CO.sub.2R.sup.ee, —OC(=Q)R.sup.ee, —OCO.sub.2R.sup.ee, —C(═O)N(R.sup.ff).sub.2, —OC(═O)N(R.sup.ff).sub.2, —NR.sup.ffC(═O)R.sup.ee, —NR.sup.ffCO.sub.2R.sup.ee, —NR.sup.ffC(═O)N(R.sup.ff).sub.2, —C(═NR.sup.ff)OR.sup.ee, —OC(═NR.sup.ff)R.sup.ee, —OC(═NR.sup.ff)OR.sup.ee, —C(═NR.sup.ff)N(R.sup.ff).sub.2, —OC(═NR.sup.ff)N(R.sup.ff).sub.2, —NR.sup.ffC(═NR.sup.ff)N(R.sup.ff).sub.2, —NR.sup.ffSO.sub.2R.sup.ee, —SO.sub.2N(R.sup.ff).sub.2, —SO.sub.2R.sup.ee, —SO.sub.2OR.sup.ee, —OSO.sub.2R.sup.ee, —S(═O)R.sup.ee, —Si(R.sup.ee).sub.3, —OSi(R.sup.ee).sub.3, —C(═S)N(R.sup.ff).sub.2, —C(═O)SR.sup.ee, —C(═S)SR.sup.ee, —SC(═S)SR.sup.ee, —P(═O).sub.2R.sup.ee, —P(═O)(R.sup.ee).sub.2, —OP(═O)(R.sup.ee).sub.2, —OP(═O)(OR.sup.ee).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, heteroC.sub.1-6alkyl, heteroC.sub.1-6alkenyl, heteroC.sub.2-6alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl, 5-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups, or two geminal R.sup.dd substituents can be joined to form ═O or ═S;
(205) each instance of R.sup.ee is, independently, selected from C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, heteroC.sub.1-6 alkyl, heteroC.sub.2-6alkenyl, heteroC.sub.2-6 alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups;
(206) each instance of R.sup.ff is, independently, selected from hydrogen, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, heteroC.sub.1-6alkyl, heteroC.sub.2-6alkenyl, heteroC.sub.2-6alkynyl, C.sub.3-10 carbocyclyl, 3-10 membered heterocyclyl, C.sub.6-10 aryl and 5-10 membered heteroaryl, or two R.sup.ff groups are joined to form a 3-10 membered heterocyclyl or 5-10 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R.sup.gg groups; and
(207) each instance of R.sup.gg is, independently, halogen, —CN, —NO.sub.2, —N.sub.3, —SO.sub.2H, —SO.sub.3H, —OH, —OC.sub.1-6 alkyl, —ON(C.sub.1-6 alkyl).sub.2, —N(C.sub.1-6 alkyl).sub.2, —N(C.sub.1-6 alkyl).sub.3.sup.+X.sup.− —NH(C.sub.1-6 alkyl).sub.2.sup.+X.sup.−, —NH.sub.2(C.sub.1-6 alkyl).sup.+X.sup.−, —NH.sub.3.sup.+X.sup.−, —N(OC.sub.1-6 alkyl)(C.sub.1-6 alkyl), —N(OH)(C.sub.1-6 alkyl), —NH(OH), —SH, —SC.sub.1-6 alkyl, —SS(C.sub.1-6 alkyl), —C(═O)(C.sub.1-6 alkyl), —CO.sub.2H, —CO.sub.2(C.sub.1-6 alkyl), —OC(═O)(C.sub.1-6 alkyl), —OCO.sub.2(C.sub.1-6 alkyl), —C(═O)NH.sub.2, —C(═O)N(C.sub.1-6 alkyl).sub.2, —OC(═O)NH(C.sub.1-6 alkyl), —NHC(═O)(C.sub.1-6 alkyl), —N(C.sub.1-6 alkyl)C(═O)(CM alkyl), —NHCO.sub.2(C.sub.1-6 alkyl), —NHC(═O)N(C.sub.1-6 alkyl).sub.2, —NHC(═O)NH(C.sub.1-6 alkyl), —NHC(═O)NH.sub.2, —C(═NH)O(C.sub.1-6 alkyl), —OC(═NH)(C.sub.1-6 alkyl), —OC(═NH)OC.sub.1-6 alkyl, —C(═NH)N(C.sub.1-6 alkyl).sub.2, —C(═NH)NH(C.sub.1-6 alkyl), —C(═NH)NH.sub.2, —OC(═NH)N(C.sub.1-6 alkyl).sub.2, —OC(NH)NH(C.sub.1-6 alkyl), —OC(NH)NH.sub.2, —NHC(NH)N(C.sub.1-6 alkyl).sub.2, —NHC(═NH)NH.sub.2, —NHSO.sub.2(C.sub.1-6 alkyl), —SO.sub.2N(C.sub.1-6 alkyl).sub.2, —SO.sub.2NH(C.sub.1-6 alkyl), —SO.sub.2NH.sub.2, —SO.sub.2C.sub.1-6 alkyl, —SO.sub.2OC.sub.1-6 alkyl, —OSO.sub.2C.sub.1-6 alkyl, —SOC.sub.1-6 alkyl, —Si(C.sub.1-6 alkyl).sub.3, —OSi(C.sub.1-6 alkyl).sub.3-C(═S)N(C.sub.1-6 alkyl).sub.2, C(═S)NH(C.sub.1-6 alkyl), C(═S)NH.sub.2, —C(═O)S(C.sub.1-6 alkyl), —C(═S)SC.sub.1-6 alkyl, —SC(═S)SC.sub.1-6 alkyl, —P(═O).sub.2(C.sub.1-6 alkyl), —P(═O)(C.sub.1-6 alkyl).sub.2, —OP(═O)(C.sub.1-6 alkyl).sub.2, —OP(═O)(OC.sub.1-6 alkyl).sub.2, C.sub.1-6 alkyl, C.sub.1-6 perhaloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, heteroC.sub.1-6alkyl, heteroC.sub.2-6 alkenyl, heteroC.sub.2-6alkynyl, C.sub.3-10 carbocyclyl, C.sub.6-10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two geminal R.sup.gg substituents can be joined to form ═O or ═S; wherein X.sup.− is a counterion.
(208) In the claims, as well as in the specification above, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “holding,” and the like are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases “consisting of” and “consisting essentially of” shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.