Biosyntheses of salinosporamide A and its analogs and related methods of making salinosporamide A and its analogs
Disclosed are methods of modulating biosynthesis of Salinosporamide A and its analogs, which are useful in treating cancer, inflammatory conditions, and/or infectious disease. The methods involve, for example, genetic manipulation, selection of reagents in the fermentation feedstock, and selection of fermentation conditions.
1. A method of producing a first compound of Formula I,
wherein the dashed lines represent a single or a double bond, wherein R 1 is separately selected from the group consisting of a hydrogen, a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, where n is equal to 1 or 2, and if n is equal to 2, then R 1 can be the same or different;
wherein R 2 is selected from the group consisting of hydrogen, a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl;
wherein R 3 is selected from the group consisting of a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl;
wherein each of E 1 , E 2 , E 3 and E 4 is a substituted or unsubstituted heteroatom;
the method comprising culturing a strain of Salinospora in a saline medium using a feedstock that is deficient in or has an excess of one or more of proprionate, butyrate, or valerate compared to levels of proprionate, butyrate, and valerate found in a medium comprising starch, Hy soy, and yeast in a ratio of 10 g to 4 g to 4 g such that the relative yield of the first compound in comparison to the yield of a second compound of Formula I is substantially different than the relative yield of the first compound in comparison to the yield of the second compound obtained when the strain is cultured in a medium comprising starch, Hy soy, and yeast in a ratio of 10 g to 4 g to 4 g.
2. The method of claim 1 , wherein the first compound has the structure of Formula II:
wherein each R 4 is separately selected from the group consisting of a hydrogen, a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, where m is equal to 1 or 2, and if m is equal to 2, then R 4 can be the same or different; and
wherein E 5 is a substituted or unsubstituted heteroatom.
3. The method of claim 1 , wherein the first compound has the structure of Formula III:
wherein each R 4 is separately selected from the group consisting of a hydrogen, a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, where m is equal to 1 or 2, and if m is equal to 2, then R 4 can be the same or different; and
wherein E 5 is a substituted or unsubstituted heteroatom.
4. The method of claim 1 , wherein the first compound has the structure of Formula IV:
wherein R 5 is separately selected from the group consisting of a hydrogen, a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, where m is equal to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11, and if m is greater than 1, then R 5 can be the same or different; and
wherein E 5 is a substituted or unsubstituted heteroatom.
5. The method of claim 1 , wherein the first compound has the structure of Formula V:
wherein R 5 is separately selected from the group consisting of a hydrogen, a halogen, mono-substituted, poly-substituted or unsubstituted variants of the following residues: saturated C 1 -C 24 alkyl, unsaturated C 2 -C 24 alkenyl or C 2 -C 24 alkynyl, acyl, acyloxy, alkyloxycarbonyloxy, aryloxycarbonyloxy, cycloalkyl, cycloalkenyl, alkoxy, cycloalkoxy, aryl, heteroaryl, arylalkoxy carbonyl, alkoxy carbonylacyl, amino, aminocarbonyl, aminocarboyloxy, nitro, azido, phenyl, cycloalkylacyl, hydroxy, alkylthio, arylthio, oxysulfonyl, carboxy, cyano, and halogenated alkyl including polyhalogenated alkyl, where m is equal to 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11, and if m is greater than 1, then R 5 can be the same or different; and
wherein E 5 is a substituted or unsubstituted heteroatom.
6. The method of claim 1 , wherein the first compound is the compound of Formula II-16:
7. The method of claim 1 , wherein the other second compound is the compound of Formula II-17:
8. The method of claim 1 , wherein said feedstock is deficient in one or more reagent selected from proprionate, butyrate, and valerate.
9. The method of claim 8 , wherein the reagent is proprionate.
10. The method of claim 8 , wherein the reagent is butyrate.
11. The method of claim 8 , wherein the reagent is valerate.
12. The method of claim 1 , wherein said feedstock comprises an excess of one or more reagent selected from proprionate, butyrate, and valerate.
13. The method of claim 12 , wherein the reagent is proprionate.
14. The method of claim 12 , wherein the reagent is butyrate.
15. The method of claim 12 , wherein the reagent is valerate.
16. The method of claim 1 , wherein said feedstock is deficient in butyrate,
wherein the first compound is the compound of Formula II-16:
and wherein the second compound is the compound of Formula II-17:
17. The method of claim 1 , wherein said feedstock comprises an excess of propionate,
wherein the first compound is the compound of Formula II-20 or the compound of Formula II-29:
18. The method of claim 1 , wherein said feedstock comprises an excess of valerate,
wherein the first compound is the compound of Formula II-30: