US20140080175A1 - Processes for preparing tubulysin derivatives and conjugates thereof - Google Patents

Processes for preparing tubulysin derivatives and conjugates thereof Download PDF

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US20140080175A1
US20140080175A1 US13/841,349 US201313841349A US2014080175A1 US 20140080175 A1 US20140080175 A1 US 20140080175A1 US 201313841349 A US201313841349 A US 201313841349A US 2014080175 A1 US2014080175 A1 US 2014080175A1
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compound
formula
optionally substituted
alkyl
cycloalkyl
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Iontcho R. Vlahov
Hari Krishna R. Santhapuram
Paul J. Kleindl
Christopher P. Leamon
Fei You
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Endocyte Inc
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Endocyte Inc
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C241/00—Preparation of compounds containing chains of nitrogen atoms singly-bound to each other, e.g. hydrazines, triazanes
    • C07C241/02—Preparation of hydrazines
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/62—Oxygen or sulfur atoms
    • C07D213/70—Sulfur atoms
    • C07D213/71—Sulfur atoms to which a second hetero atom is attached
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/72—Nitrogen atoms
    • C07D213/76—Nitrogen atoms to which a second hetero atom is attached
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
    • C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
    • C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D277/56—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
    • C07F7/02—Silicon compounds
    • C07F7/08—Compounds having one or more C—Si linkages
    • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
    • C07F7/1804—Compounds having Si-O-C linkages
    • C07F7/1872—Preparation; Treatments not provided for in C07F7/20
    • C07F7/188—Preparation; Treatments not provided for in C07F7/20 by reactions involving the formation of Si-O linkages
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
    • C07F7/02—Silicon compounds
    • C07F7/08—Compounds having one or more C—Si linkages
    • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
    • C07F7/1804—Compounds having Si-O-C linkages
    • C07F7/1872—Preparation; Treatments not provided for in C07F7/20
    • C07F7/1892—Preparation; Treatments not provided for in C07F7/20 by reactions not provided for in C07F7/1876 - C07F7/1888
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/02—Linear peptides containing at least one abnormal peptide link

Definitions

  • the invention described herein pertains to processes for preparing tubulysin derivatives, conjugates of tubulysins, and intermediates therefore.
  • the tubulysins are members of a new class of natural products isolated from myxobacterial species (F. Sasse, et al., J. Antibiot. 2000, 53, 879-885). As cytoskeleton interacting agents, the tubulysins are mitotic poisons that inhibit tubulin polymerization and lead to cell cycle arrest and apoptosis (H. Steinmetz, et al., Chem. Int. Ed. 2004, 43, 4888-4892; M. Khalil, et al., ChemBioChem. 2006, 7, 678-683; G. Kaur, et al., Biochem. J. 2006, 396, 235-242).
  • Tubulysins are extremely potent cytotoxic molecules, and exceed the cell growth inhibition of many other clinically relevant traditional chemotherapeutics, including epothilones, paclitaxel, and vinblastine. Furthermore, they are potent against multidrug resistant cell lines (A. Dömling, et al., Mol. Diversity. 2005, 9, 141-147). These compounds show high cytotoxicity tested against a panel of cancer cell lines with IC 50 values in the low picomolar range; thus, they are of interest as potential anticancer therapeutics. However, tubulysins have been reported to exhibit a narrow, or in some cases nonexistent, therapeutic window such that disease treatment with tubulysins is hampered by toxicity and other unwanted side effects.
  • tubulysins have been conjugated with targeting agents to improve their therapeutic window.
  • R A OH
  • tubulysin derivatives tubulysin analogs, and other tubulysin conjugate intermediates that are useful for preparing such targeted conjugates.
  • tubulysin linker derivatives useful for preparing vitamin receptor binding tubulysin conjugates (also referred to herein as tubulysin linker derivatives) are described herein.
  • tubulysin linker derivatives include linear tetrapeptoid backbones, including illustrative compounds having the following formula
  • Ar 1 is optionally substituted aryl or optionally substituted heteroaryl
  • Ar 2 is optionally substituted aryl or optionally substituted heteroaryl
  • L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents hydrogen, or 1 to 4 substituents each independently selected from the group consisting of amino or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof, and alkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, and heteroarylheteroalkyl, each of which is optionally substituted;
  • Y is R 2 C(O)O or R 12 O;
  • R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 4 is optionally substituted alkyl or optionally substituted cycloalkyl
  • R 3 is optionally substituted alkyl
  • R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 7 is optionally substituted alkyl
  • R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted;
  • R Ar represents hydrogen, or 1 to 4 substituents each independently selected from the group consisting of amino or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof, and alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, and heteroarylheteroalkyl; and n is 1, 2, 3, or 4.
  • tubulysin linker derivative has formula T1
  • tubulysin linker derivative has formula T2
  • Ar 2 is optionally substituted aryl.
  • Ar 2 is optionally substituted heteroaryl.
  • tubulysins described herein are more particularly comprised of one or more non-naturally occurring or hydrophobic amino acid segments, such as N-methyl pipecolic acid (Mep), isoleucine (Ile),
  • tubuvaline includes compounds of the following formula,
  • R 3 and Ar 1 are as described in any of the embodiment described herein.
  • a common feature in the molecular architecture of potent natural occurring tubulysins is the acid and/or base sensitive N-acyloxymethyl substituent (or a N,O-acetal of formaldehyde) represented by R 2 CO 2 CH 2 in the formula (T1).
  • the compounds described herein are NHNH—C(O)O-L-SS-Ar 2 derivatives of naturally occurring tubulysins.
  • An illustrative group of tubulysin derivatives described herein are those having formula 1.
  • tubulysins conjugated to vitamin receptor binding moieties for targeted and/or selective delivery to cell populations expressing, overexpressing or selectively expressing cell surface vitamin receptors necessitates further modification of the highly toxic tubulysins.
  • Described herein are improved processes for making natural tubulysins analogs or derivatives, which are useful for preparing vitamin receptor binding tubulysin conjugates including compounds of formula (T) and formula (I).
  • Vitamin receptor binding conjugates of tubulysins are described in U.S. Patent Publication 2010/0048490, the disclosure of which is incorporated herein by reference in its entirety.
  • processes for derivatives or analogs of natural tubulysins including compounds of formula (T).
  • vitamin receptor binding conjugates of tubulysins are described. The processes include one or more steps described herein.
  • a process for preparing a compound of formula B, wherein R 5 and R 6 are as described in the various embodiments herein, such as each being independently selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 8 is C1-C6 n-alkyl; wherein the process comprises the step of treating a compound of formula A with a silylating agent, such as triethylsilyl chloride, and a base, such as imidazole in an aprotic solvent.
  • a silylating agent such as triethylsilyl chloride
  • R 5 and R 6 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula C, wherein R 5 and R 6 are as described in the various embodiments herein, such as each being independently selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 8 is C1-C6 n-alkyl; and R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; wherein the process comprises the step of treating a compound of formula B with a base and a compound of the formula ClCH 2 OC(O)R 2 in an aprotic solvent at a temperature below ambient temperature, such as in the range from about ⁇ 78° C. to about 0° C.; wherein the molar ratio of the compound of the formula ClCH 2 OC(O)R 2 to the compound of formula B is from about 1 to about 1.5.
  • R 2 , R 5 and R 6 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula D, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 8 is C1-C6 n-alkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the steps of
  • R 2 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula F, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating compound D with a hydrolase enzyme.
  • a process for preparing a compound of formula F, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating compound D with a trialkyltin hydroxide (e.g. trimethyltin hydroxide).
  • R 2 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula AF, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of contacting compound D with an alcohol, R 12 OH, where R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and a transesterification catalyst.
  • the transesterification catalyst is trifluoroacetic acid (TFA).
  • the transesterification catalyst is selected from the group consisting of (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the transesterification catalyst is(R 13 ) 2 SnO.
  • R 13 include methyl, n-butyl, n-octyl, phenyl, o-MeO-phenyl, p-MeO phenyl, phenethyl, and benzyl.
  • R 5 , R 6 , R 12 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula G, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating the silyl ether of compound F with a non-basic fluoride containing reagent.
  • R 2 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula AG, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of contacting compound F with an alcohol, R 12 OH, where R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and a transesterification catalyst.
  • the transesterification catalyst is trifluoroacetic acid (TFA).
  • the transesterification catalyst is selected from the group consisting of (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the transesterification catalyst is (R 13 ) 2 SnO.
  • R 13 is methyl, n-butyl, n-octyl, phenyl, o-MeO-phenyl, p-MeO phenyl, phenethyl, and benzyl.
  • R 2 , R 5 , R 6 , R 7 , and R 12 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula BG, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 12 is as described in the various embodiments herein, such as being selected from alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and R 2 is optionally substituted alkyl; wherein the process comprises the step of contacting compound AF with a metal hydroxide or carbonate.
  • Illustrative examples of a metal hydroxide or carbonate include LiOH, Li 2 CO 3 , NaOH, Na 2 CO 3 , KOH, K 2 CO 3 , Ca(OH) 2 , CaCO 3 , Mg(OH) 2 , MgCO 3 , and the like.
  • R 5 , R 6 , R 7 , and R 12 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula H, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating a compound of formula G with an acylating agent of formula R 4 C(O)X 2 , where X 2 is a leaving group.
  • R 2 , R 4 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula AH, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 12 is as described in the various embodiments herein, such as being selected from alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating a compound of formula BG with an acylating agent of formula R 4 C(O)X 2 , where X 2 is a leaving group.
  • R 4 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a tubulysin linker derivative of formula (T1), wherein Ar 1 is optionally substituted aryl; Ar 2 is optionally substituted aryl or optionally substituted heteroaryl; L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 1 is hydrogen, optionally substituted alkyl, optionally substituted arylalkyl or a pro drug forming group;
  • R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl;
  • R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl
  • the process comprises the step of forming an active ester intermediate from a compound of formula H; and reacting the active ester intermediate with a compound of the formula I to give a compound of the formula T.
  • Ar 1 , Ar 2 , R 1 , R 2 , R 4 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a tubulysin linker derivative of formula (T2), wherein Ar 1 is optionally substituted aryl; Ar 2 is optionally substituted aryl or optionally substituted heteroaryl; L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 1 is hydrogen, optionally substituted alkyl, optionally substituted arylalkyl or a pro-drug forming group;
  • R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl;
  • R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and
  • R 7 is optionally substituted alkyl; wherein the process comprises the step of contacting compound T, with an alcohol, R 12 OH, where R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and a transesterification catalyst.
  • the transesterification catalyst is trifluoroacetic acid (TFA).
  • the transesterification catalyst is selected from the group consisting of (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the transesterification catalyst is (R 13 ) 2 SnO.
  • R 13 are methyl, n-butyl, n-octyl, phenyl, o-MeO-phenyl, p-MeO phenyl, phenethyl, and benzyl.
  • Ar 1 , Ar 2 , R 1 , R 2 , R 4 , R 5 , R 6 , R 7 , and R 12 may each include conventional protection groups on the optional substituents.
  • a process for preparing a tubulysin linker derivative of formula (T2), wherein Ar 1 is optionally substituted aryl; Ar 2 is optionally substituted aryl or optionally substituted heteroaryl; L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 1 is hydrogen, optionally substituted alkyl, optionally substituted arylalkyl or a pro-drug forming group;
  • R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl;
  • R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and
  • R 7 is optionally substituted alkyl;
  • the process comprises the step of forming an active ester intermediate from a compound of formula AH; and reacting the active ester intermediate with a compound of the formula I to give a compound of the formula AT.
  • Ar 1 , Ar 2 , R 1 , R 12 , R 3 , R 4 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula B, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 8 is C1-C6 n-alkyl; wherein the process comprises the step of treating a compound of formula A with triethylsilyl chloride and imidazole in an aprotic solvent.
  • the use of the less reactive TESCl may also be advantageous when the process is performed on larger scales, where higher reactivity reagents may represent a safety issue. It has also been discovered that the use of TESCl in nearly stoichiometric amounts renders the chromatographic purification step unnecessary.
  • the process is performed without subsequent purification.
  • R 5 is isopropyl.
  • R 6 is sec-butyl.
  • R 8 is methyl.
  • the silyl ether is TES.
  • the compound 4 is not purified by chromatography.
  • a process for preparing a compound of formula C, wherein R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; R 8 is C1-C6 n-alkyl; and R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; wherein the process comprises the step of treating a compound of formula B with from about 1 equivalent to about 1.5 equivalent of base and from about 1 equivalent to about 1.5 equivalent of a compound of the formula ClCH 2 OC(O)R 2 in an aprotic solvent at a temperature from about ⁇ 78° C. to about 0° C.
  • the ether analog of C′ can be used to prepare the tubulysins, the tubulysin conjugates, and the tubulysin linker compounds described herein.
  • the process of any one of the preceding embodiments is described wherein about 1 equivalent to about 1.3 equivalent of a compound of the formula ClCH 2 OC(O)R 2 is used. In another illustrative example, the process of any one of the preceding embodiments is described, wherein about 1.2 equivalent of a compound of the formula ClCH 2 OC(O)R 2 is used. In another illustrative example, the process of any one of the preceding embodiments is described wherein R 2 is n-propyl. In another alternative of the foregoing embodiments, and each additional embodiment described herein, R 2 is CH 2 CH(CH 3 ) 2 , CH 2 CH 2 CH 3 , CH 2 CH 3 , CH ⁇ C(CH 3 ) 2 , or CH 3 .
  • N,O-acetal 5 a process for preparing the N,O-acetal 5 is described.
  • compound 2 is treated with 1.1 equivalent of potassium hexamethyldisilazane (KHMDS) and 1.2 equivalent of chloromethyl butanoate in a nonprotic solvent at about ⁇ 45° C.
  • KHMDS potassium hexamethyldisilazane
  • chloromethyl butanoate in a nonprotic solvent at about ⁇ 45° C.
  • the product formed by any of the preceding examples may be used without chromatographic purification.
  • the ether analogs of compound 5 can also be used to prepare the tubulysins, tubulysin linker compounds
  • a process for preparing a compound of formula D, wherein R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and cycloalkyl; R 8 is C1-C6 n-alkyl; R 2 is selected from the group consisting of optionally substituted alkyl and cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the steps of
  • a mixture of compound 5 and the pentafluorophenyl ester of D-N-methyl-pipecolic acid is reduced using H 2 and a palladium-on-charcoal catalyst (Pd/C) to yield compound 6.
  • Pd/C palladium-on-charcoal catalyst
  • n is 3.
  • R 7 is methyl.
  • the ether analogs of 5 and 6 can be used to prepare the tubulysins, tubulysin conjugates, and tubulysin linker compounds described herein. It has been previously reported that removal of the protecting group from the secondary hydroxyl group leads to an inseparable mixture of the desired product 5a and a cyclic O,N-acetal side-product, 5b, as shown in the following scheme.
  • iso-7a results from rearrangement of the butyrate group from the N-hydroxymethyl group to the secondary hydroxyl group, as shown below.
  • a process for preparing a compound of formula AF, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of contacting compound D with an alcohol, R 12 OH, where R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and a transesterification catalyst.
  • the transesterification catalyst is trifluoroacetic acid (TFA).
  • the transesterification catalyst is selected from the group consisting of (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the catalyst is (R 13 ) 2 SnO.
  • R 13 is methyl, n-butyl, n-octyl, phenyl, o-MeO-phenyl, p-MeO phenyl, phenethyl, and benzyl.
  • R 5 , R 6 , R 12 , and R 7 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula F, wherein R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating compound D with a hydrolase enzyme.
  • the preceding process wherein the treating step comprises adding a solution of compound D in a water miscible solvent to a buffered solution containing the hydrolase enzyme at a rate which minimizes precipitation of the ester.
  • the ester is added over a period of from about 24 hours to about 100 hours.
  • the ester is added over a period of from about 48 hours to about 100 hours.
  • R 8 is methyl.
  • the embodiment of any one of the preceding embodiments wherein the hydrolase enzyme is an esterase is described.
  • the embodiment of any of the preceding embodiments wherein the esterase is a pig liver esterase is described.
  • a process for preparing a compound of formula F, wherein R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating compound D with a trialkyltin hydroxide.
  • the trialkyltin hydroxide is trimethyltin hydroxide.
  • a solution of compound 4 in dimethyl sulfoxide (DMSO) is added over a period of 90 hours, to a buffered solution of pig liver esterase.
  • the buffer is a phosphate buffer.
  • the solution of the enzyme has a pH of 6.5 to 8.5.
  • the solution of the enzyme has a pH of 7.4 to 7.8. It is appreciated that the buffering material used can be any buffer compatible with the hydrolase enzyme used to remove the ester.
  • Tripeptide methyl ester 6 (reported in an earlier patent application [1]) was treated with trimethyltin hydroxide to yield corresponding acid 7.
  • the triethylsilyl group was removed by treatment with hydrogen flouride-triethyl amine complex, and acetylated, resulting in acetyl-tripeptide acid 8.
  • a process for preparing a compound of formula G, wherein R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating the silyl ether of compound F with a non basic fluoride reagent. It has been discovered herein that use of basic conditions can lead to the production of a by-product arising from the rearrangement of the ester group to give compound G′.
  • compound 7 is treated with Et 3 N.3HF to cleave the TES-ether in the preparation of the corresponding alcohol 6′.
  • Et 3 N.3HF a non-basic fluoride reagent to cleave the silyl ether of compounds F
  • other non-basic fluoride reagents to cleave the silyl ether of compounds F may be used in the methods and processes described herein, including but not limited to pyridine.HF, and the like to cleave the TES-ether.
  • a process for preparing a compound of formula AG, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of contacting compound F with an alcohol, R 12 OH, where R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and a transesterification catalyst.
  • the transesterification catalyst is trifluoroacetic acid (TFA).
  • the transesterification catalyst is selected from the group consisting of (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the catalyst is (R 13 ) 2 SnO.
  • R 13 is methyl, n-butyl, n-octyl, phenyl, o-MeO-phenyl, p-MeO phenyl, phenethyl, and benzyl.
  • R 2 , R 5 , R 6 , R 2 , and R 12 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula BG, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 is as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 12 is as described in the various embodiments herein, such as being selected from alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and R 2 is optionally substituted alkyl; wherein the process comprises the step of contacting compound AF with a metal hydroxide or carbonate.
  • Illustrative examples of a metal hydroxide or carbonate include LiOH, Li 2 CO 3 , NaOH, Na 2 CO 3 , KOH, K 2 CO 3 , Ca(OH) 2 , CaCO 3 , Mg(OH) 2 , MgCO 3 , and the like.
  • R 5 , R 6 , R 7 , and R 12 may each include conventional protection groups on the optional substituents.
  • a process for preparing a compound of formula H, wherein R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; R 2 and R 4 are independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; and R 7 is optionally substituted alkyl; wherein the process comprises the step of treating a compound of formula G with an acylating agent of formula R 4 C(O)X 2 , where X 2 is a leaving group. It is appreciated that the resulting product may contain varying amounts of the mixed anhydride of compound H and R 4 CO 2 H.
  • the process described in the preceding embodiment further comprises the step of treating the reaction product with water to prepare H, free of or substantially free of anhydride.
  • the process of the preceding embodiments wherein X 2 is R 4 CO 2 is described.
  • the process of any one of the preceding embodiments wherein R 4 is C1-C4 alkyl is described.
  • R 4 is methyl.
  • the process of any one of the preceding embodiments wherein R 6 is sec-butyl is described.
  • the process of any one of the preceding embodiments wherein R 7 is methyl is described.
  • the process of any one of the preceding embodiments wherein R 5 is iso-propyl is described.
  • compound 6′ is treated with acetic anhydride in pyridine. It has been discovered herein that shortening the time for this step of the process improves the yield of compound H by limiting the amount of the previously undescribed alternative acylation side products, such as formula 7a that are formed. It is appreciated that the resulting product may contain varying amounts of the mixed anhydride of 8 and acetic acid.
  • treatment of the reaction product resulting from the preceding step with water in dioxane yields compound 8, free of or substantially free of anhydride. It is to be understood that other solvents can be substituted for dioxane in the hydrolysis of the intermediate mixed anhydride. Alternatively, the step may be performed without solvent.
  • a process for preparing a compound of formula AH, wherein R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; R 12 is as described in the various embodiments herein, such as being selected from alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and R 2 is optionally substituted alkyl; wherein the process comprises the step of treating a compound of formula BG with an acylating agent of formula R 4 C(O)X 2 , where X 2 is a leaving group.
  • R 4 , R 5 , R 6 , and R 2 may each include conventional protection groups on the optional substituents.
  • mixed carbonate 11 was prepared from 4-nitropyridyldisulfide ethanol 12 and 4-nitrophenyl chloroformate as shown.
  • Boc-Tut-hydrazide 13 was prepared from corresponding acid 10 and coupled with mixed carbonate 11 to yield activated Boc-Tut fragment 14.
  • a process for preparing a tubulysin linker derivative T, wherein An is optionally substituted aryl; Ar 2 is optionally substituted aryl or optionally substituted heteroaryl; L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl;
  • R 2 and R 4 are independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl; and
  • R 7 is optionally substituted alkyl;
  • compound H is treated with an excess amount of active ester forming agent and pentafluorophenol to form the pentafluorophenol ester of compound H, followed by removal of the excess active ester forming agent prior to the addition of compound I.
  • Ar 1 is phenyl.
  • Ar 1 is substituted phenyl.
  • Ar 1 is 4-substituted phenyl.
  • Ar 1 is R A -substituted phenyl.
  • Ar 1 is 4-hydroxyphenyl, or a hydroxyl protected form thereof.
  • Ar 2 is substituted pyridyl.
  • Ar 2 is substituted 2-pyridyl.
  • Ar 2 is 3-nitro-2-pyridyl.
  • R 3 is methyl.
  • compound 8 is treated with an excess amount of a polymeric version of a carbodiimide and pentafluorophenol to form the pentafluorophenyl ester of 8, the polymeric carbodiimide is removed by filtration; and compound 9 is added to the solution to yield tubulysin B linker derivative 2.
  • the process of any one of the preceding embodiments wherein the polymeric carbodiimide is polystyrene-CH 2 —N ⁇ C ⁇ N-cyclohexane (PS-DCC) is described.
  • the ether analog of compound 8 can be converted to the ether analog of compound 2, via the ether analog of compound 15, where R is allyl, or CH 2 (CH 2 ) n CH 3 , and n is 1, 2, 3, 4, 5, or 6.
  • a process for preparing a tubulysin linker derivative of formula (T2), wherein Ar 1 is optionally substituted aryl; Ar 2 is optionally substituted aryl or optionally substituted heteroaryl; L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • Ra, Rb, and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 1 is hydrogen, optionally substituted alkyl, optionally substituted arylalkyl or a pro-drug forming group;
  • R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl;
  • R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and
  • R 7 is optionally substituted alkyl;
  • the process comprises the step of contacting compound T, with an alcohol, R 12 OH, where R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted; and a transesterification catalyst.
  • the transesterification catalyst is trifluoroacetic acid (TFA).
  • the transesterification catalyst is selected from the group consisting of (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the catalyst is (R 13 ) 2 SnO.
  • R 13 are methyl, n-butyl, n-octyl, phenyl, o-MeO-phenyl, p-MeO-phenyl, phenethyl, and benzyl. It is to be understood that Ar 1 , Ar 2 , R 1 , R 2 , R 4 , R 5 , R 6 , R 7 , and R 12 may each include conventional protection groups on the optional substituents.
  • a process for preparing a tubulysin linker derivative of formula (T2), wherein Ar 1 is optionally substituted aryl; Ar 2 is optionally substituted heteroaryl; L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • Ra, Rb, and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 1 is hydrogen, optionally substituted alkyl, optionally substituted arylalkyl or a pro-drug forming group;
  • R 5 and R 6 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl;
  • R 2 and R 4 are as described in the various embodiments herein, such as being selected from optionally substituted alkyl or optionally substituted cycloalkyl; and
  • R 7 is optionally substituted alkyl;
  • the process comprises the step of forming an active ester intermediate from a compound of formula AH; and reacting the active ester intermediate with a compound of the formula Ito give a compound of the formula AT.
  • Ar 1 , Ar 2 , R 1 , R 12 , R 3 , R 4 , R 5 , R 6 , and R 7 may each include conventional protection groups on the optional substituents.
  • a compound having formula D wherein the compound is free of or substantially free of a compound having formula C-1 is described, where in R 2 , R 5 , R 6 , R 7 , and R 8 are as described in any of the embodiments described herein. Without being bound by theory, it is believed herein that compounds C-1 are formed from the corresponding compounds C via an acyl transfer.
  • compound 4 free of or substantially free of compound 8a and/or compound iso-6 is described.
  • an optically pure form of compound 6 is formed.
  • a compound G where the compound is free of or substantially free of a compound G′ is described, wherein R 2 , R 5 , R 6 , and R 7 are as described in any of the embodiments described herein.
  • compound 6′ is described, wherein compound 6′ is free of or substantially free of the isomer of G′ shown below
  • compound 8 is described, wherein compound 8 is free of or substantially free of compound 8b is described
  • R a , R b , and R are each independently selected from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring.
  • R a and R b are hydrogen.
  • R a and R b are hydrogen.
  • p is 1.
  • R a and R b are hydrogen and p is 1.
  • a compound H is described wherein R 4 is Me and R 2 , R 5 , R 6 , and R 2 are as described in any of the embodiments described herein; and the compound H is free of or substantially free of the compound H wherein R 4 and R 2 are both Me.
  • R 5 is isopropyl
  • R 6 is sec-butyl
  • R 8 is methyl
  • R 2 is CH 2 CH(CH 3 ) 2 , CH 2 CH 2 CH 3 , CH 2 CH 3 , CH ⁇ C(CH 3 ) 2 , or CH 3 .
  • n 3.
  • R 7 is methyl
  • R 8 is methyl
  • R 4 is methyl
  • Ar 1 is phenyl. In another alternative of the foregoing embodiments, and each additional embodiment described herein, Ar 1 is substituted phenyl. In another alternative of the foregoing embodiments, and each additional embodiment described herein, Ar 1 is 4-substituted phenyl. In another alternative of the foregoing embodiments, and each additional embodiment described herein, Ar 1 is R A -substituted phenyl. In another alternative of the foregoing embodiments, and each additional embodiment described herein, Ar 1 is 4-hydroxyphenyl, or a hydroxyl protected form thereof.
  • R 3 is methyl
  • Ar 1 is optionally substituted aryl or optionally substituted heteroaryl
  • Ar 2 is optionally substituted aryl or optionally substituted heteroaryl
  • L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • Y is acyloxy or R 12 O
  • R 3 is optionally substituted alkyl
  • R 4 is optionally substituted alkyl or optionally substituted cycloalkyl
  • R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 7 is optionally substituted alkyl
  • R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted;
  • n 1, 2, 3, or 4;
  • Ar 1 is optionally substituted aryl or optionally substituted heteroaryl
  • Ar 2 is optionally substituted aryl or optionally substituted heteroaryl
  • L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 3 is optionally substituted alkyl
  • R 4 is optionally substituted alkyl or optionally substituted cycloalkyl
  • R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 2 is optionally substituted alkyl
  • n 1, 2, 3, or 4;
  • Ar 1 is optionally substituted aryl
  • Ar 2 is optionally substituted aryl or optionally substituted heteroaryl
  • L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 2 is selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 12 is alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, arylalkyl or heteroarylalkyl, each of which is optionally substituted;
  • R 3 is optionally substituted alkyl
  • R 4 is optionally substituted alkyl or optionally substituted cycloalkyl
  • R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 7 is optionally substituted alkyl
  • n 1, 2, 3, or 4;
  • transesterification reagent selected from TFA or (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, or (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted or
  • Ar 1 is optionally substituted aryl or optionally substituted heteroaryl
  • Ar 2 is optionally substituted aryl or optionally substituted heteroaryl
  • L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • R 3 is optionally substituted alkyl
  • R 9 is hydrogen or an amine protecting group.
  • R 2 is CH 2 CH(CH 3 ) 2 , CH 2 CH 2 CH 3 , CH 2 CH 3 , CH ⁇ C(CH 3 ) 2 , or CH 3 .
  • transesterification catalyst is selected from (R 13 ) 8 Sn 4 O 2 (NCS) 4 , (R 13 ) 2 Sn(OAc) 2 , (R 13 ) 2 SnO, (R 13 ) 2 SnCl 2 , (R 13 ) 2 SnS, (R 13 ) 3 SnOH, and (R 13 ) 3 SnOSn(R 13 ) 3 , where R 13 is independently selected from alkyl, arylalkyl, aryl, or cycloalkyl, each of which is optionally substituted.
  • the alkaline salt is selected from LiOH, Li 2 CO 3 , NaOH, Na 2 CO 3 , KOH, K 2 CO 3 , Ca(OH) 2 , CaCO 3 , Mg(OH) 2 , or MgCO 3 .
  • R 2 is CH 2 CH(CH 3 ) 2 , CH 2 CH 2 CH 3 , CH 2 CH 3 , CH ⁇ C(CH 3 ) 2 , or CH 3 .
  • R 2 is CH 2 CH(CH 3 ) 2 , CH 2 CH 2 CH 3 , CH 2 CH 3 , CH ⁇ C(CH 3 ) 2 , or CH 3 .
  • Ar 1 is optionally substituted aryl
  • Ar 2 is optionally substituted aryl or optionally substituted heteroaryl
  • L is selected from the group consisting of
  • p is an integer from about 1 to about 3
  • m is an integer from about 1 to about 4, and * indicates the points of attachment;
  • R a , R b , and R are each independently selected in each instance from the group consisting of hydrogen and alkyl; or any two of R a , R b , and R are taken together with the attached carbon atom(s) to form a carbocyclic ring;
  • R Ar represents 0 to 4 substituents selected from the group consisting of amino, or derivatives thereof, hydroxy or derivatives thereof, halo, thio or derivatives thereof, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, heteroaryl, heteroarylalkyl, heteroarylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof;
  • X is hydrogen or alkyl, including C 1-6 alkyl and C 1-4 alkyl, alkenyl, including C 2-6 alkenyl and C 2-4 alkenyl, cycloalkyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, each of which is optionally substituted; or CH 2 QR 9 ; where Q is NH, O, or S; or; and R 9 is alkyl, including C 1-6 alkyl and C 1-4 alkyl, alkenyl, including C 2-6 alkenyl and C 2-4 alkenyl, cycloalkyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, each of which is optionally substituted; or R 9 is hydrogen or C(O)R 10 , where R 10 is C 1-6 alkyl, C 2-6 alkenyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, each of which is optionally substituted;
  • R 3 is optionally substituted alkyl
  • R 4 is optionally substituted alkyl or optionally substituted cycloalkyl
  • R 5 and R 6 are each independently selected from the group consisting of optionally substituted alkyl and optionally substituted cycloalkyl;
  • R 7 is optionally substituted alkyl
  • n 1, 2, 3, or 4;
  • X is CH 2 QR 9A ; where Q is NH, O, or S; and R 9A is alkyl, including C 1-6 alkyl and C 1-4 alkyl, alkenyl, including C 2-6 alkenyl and C 2-4 alkenyl, cycloalkyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, each of which is optionally substituted; or R 9A is H or C(O)R 10 , where R 10 is C 1-6 alkyl, C 2-6 alkenyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, each of which is optionally substituted.
  • one or more of the following intermediates can be used to prepare a tubulysin, tubulysin conjugate, and/or a tubulysin linker compound.
  • processes for preparing tubulysin conjugates are described herein.
  • the processes include the step of reacting compounds (2) and the corresponding tubulysins described herein that include a 3-nitropyridin-2-ylthio activating group (Ar 2 ), such as compounds of the formulae
  • the processes include acetonitrile as a reaction solvent.
  • the processes include aqueous phosphate as a reaction medium buffer. Without being bound by theory, it is believed herein that the use of acetonitrile as a solvent leads to a more rapid homogenous reaction solution. In addition, though without being bound by theory, it is believed herein that a more rapidly formed homogenous reaction medium may improve the yield and rate at which tubulysin conjugates described herein are formed.
  • the use of 3-nitropyridin-2-ylthio activating groups may lead to faster reaction times.
  • the use of 3-nitropyridin-2-ylthio activating groups may provide better chromatographic separation between the product tubulysin conjugates and the by-product 3-nitropyridin-2-ylthiol compared to conventional pyridin-2-ylthio activating groups.
  • such better chromatographic separation may allow the use of alternative purification methods.
  • the tubulysin conjugates described herein may be purified by rapid pass-through chromatography methods, such as flash C 18 chromatography, BiotageTM C 18 chromatography systems, and the like.
  • tubulysin refers both collectively and individually to the naturally occurring tubulysins, and the analogs and derivatives of tubulysins. Illustrative examples of a tubulysin are shown in Table 1.
  • tubulysin linker derivatives generally refers to the compounds described herein and analogs and derivatives thereof. It is also to be understood that in each of the foregoing, any corresponding pharmaceutically acceptable salt is also included in the illustrative embodiments described herein.
  • derivatives may include prodrugs of the compounds described herein, compounds described herein that include one or more protection or protecting groups, including compounds that are used in the preparation of other compounds described herein.
  • tubulysin linker derivatives also refers to prodrug derivatives of the compounds described herein, and including prodrugs of the various analogs and derivatives thereof.
  • tubulysin linker derivatives refers to both the amorphous as well as any and all morphological forms of each of the compounds described herein.
  • tubulysin linker derivatives refers to any and all hydrates, or other solvates, of the compounds described herein.
  • the compounds described herein may contain one or more chiral centers, or may otherwise be capable of existing as multiple stereoisomers. It is to be understood that in one embodiment, the invention described herein is not limited to any particular stereochemical requirement, and that the compounds, and compositions, methods, uses, and medicaments that include them may be optically pure, or may be any of a variety of stereoisomeric mixtures, including racemic and other mixtures of enantiomers, other mixtures of diastereomers, and the like. It is also to be understood that such mixtures of stereoisomers may include a single stereochemical configuration at one or more chiral centers, while including mixtures of stereochemical configuration at one or more other chiral centers.
  • the compounds described herein may include geometric centers, such as cis, trans, (E)-, and (Z)-double bonds. It is to be understood that in another embodiment, the invention described herein is not limited to any particular geometric isomer requirement, and that the compounds, and compositions, methods, uses, and medicaments that include them may be pure, or may be any of a variety of geometric isomer mixtures. It is also to be understood that such mixtures of geometric isomers may include a single configuration at one or more double bonds, while including mixtures of geometry at one or more other double bonds.
  • aprotic solvent refers to a solvent which does not yield a proton to the solute(s) under reaction conditions.
  • nonprotic solvents are tetrahydrofuran (THF), 2,5-dimethyl-tetrahydrofuran, 2-methyl-tetrahydrofuran, tetrahydropyran, diethyl ether, t-butyl methyl ether, dimethyl formamide, N-methylpyrrolidinone (NMP), and the like. It is appreciated that mixtures of these solvents may also be used in the processes described herein.
  • an equivalent amount of a reagent refers to the theoretical amount of the reagent necessary to transform a starting material into a desired product, i.e. if 1 mole of reagent is theoretically required to transform 1 mole of the starting material into 1 mole of product, then 1 equivalent of the reagent represents 1 mole of the reagent; if X moles of reagent are theoretically required to convert 1 mole of the starting material into 1 mole of product, then 1 equivalent of reagent represents X moles of reagent.
  • active ester forming agent generally refers to any reagent or combinations of reagents that may be used to convert a carboxylic acid into an active ester.
  • active ester generally refers to a carboxylic acid ester compound wherein the divalent oxygen portion of the ester is a leaving group resulting in an ester that is activated for reacting with compounds containing functional groups, such as amines, alcohols or sulfhydryl groups.
  • active ester-forming compounds are N-hydroxysuccinimide, N-hydroxyphthalimide, phenols substituted with electron withdrawing groups, such as but not limited to 4-nitrophenol, pentafluorophenol, N,N′-disubstituted isoureas, substituted hydroxyheteroaryls, such as but not limited to 2-pyridinols, 1-hydroxybenzotriazoles, 1-hydroxy-7-aza-benzotriazoles, cyanomethanol, and the like.
  • the reaction conditions for displacing the active ester with a compound having an amino, hydroxy or thiol group are mild.
  • reaction conditions for displacing the active ester with a compound having an amino, hydroxy or thiol group are performed at ambient or below ambient temperatures.
  • the reaction conditions for displacing the active ester with a compound having an amino, hydroxy or thiol group are performed without the addition of a strong base.
  • the reaction conditions for displacing the active ester with a compound having an amino, hydroxy or thiol group are performed with the addition of a tertiary amine base, such as a tertiary amine base having a conjugate acid pKa of about 11 or less, about 10.5 or less, and the like.
  • alkyl includes a chain of carbon atoms, which is optionally branched.
  • alkenyl and alkynyl includes a chain of carbon atoms, which is optionally branched, and includes at least one double bond or triple bond, respectively. It is to be understood that alkynyl may also include one or more double bonds. It is to be further understood that in certain embodiments, alkyl is advantageously of limited length, including C 1 -C 24 , C 1 -C 12 , C 1 -C 8 , C 1 -C 6 , and C 1 -C 4 .
  • alkenyl and/or alkynyl may each be advantageously of limited length, including C 2 -C 24 , C 2 -C 12 , C 2 -C 8 , C 2 -C 6 , and C 2 -C 4 . It is appreciated herein that shorter alkyl, alkenyl, and/or alkynyl groups may add less lipophilicity to the compound and accordingly will have different pharmacokinetic behavior.
  • Illustrative alkyl groups are, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, 2-pentyl, 3-pentyl, neopentyl, hexyl, heptyl, octyl and the like.
  • cycloalkyl includes a chain of carbon atoms, which is optionally branched, where at least a portion of the chain in cyclic. It is to be understood that cycloalkylalkyl is a subset of cycloalkyl. It is to be understood that cycloalkyl may be polycyclic. Illustrative cycloalkyl include, but are not limited to, cyclopropyl, cyclopentyl, cyclohexyl, 2-methylcyclopropyl, cyclopentyleth-2-yl, adamantyl, and the like.
  • cycloalkenyl includes a chain of carbon atoms, which is optionally branched, and includes at least one double bond, where at least a portion of the chain in cyclic. It is to be understood that the one or more double bonds may be in the cyclic portion of cycloalkenyl and/or the non-cyclic portion of cycloalkenyl. It is to be understood that cycloalkenylalkyl and cycloalkylalkenyl are each subsets of cycloalkenyl. It is to be understood that cycloalkyl may be polycyclic.
  • Illustrative cycloalkenyl include, but are not limited to, cyclopentenyl, cyclohexylethen-2-yl, cycloheptenylpropenyl, and the like. It is to be further understood that chain forming cycloalkyl and/or cycloalkenyl is advantageously of limited length, including C 3 -C 24 , C 3 -C 12 , C 3 -C 8 , C 3 -C 6 , and C 5 -C 6 . It is appreciated herein that shorter alkyl and/or alkenyl chains forming cycloalkyl and/or cycloalkenyl, respectively, may add less lipophilicity to the compound and accordingly will have different pharmacokinetic behavior.
  • heteroalkyl includes a chain of atoms that includes both carbon and at least one heteroatom, and is optionally branched.
  • Illustrative heteroatoms include nitrogen, oxygen, and sulfur. In certain variations, illustrative heteroatoms also include phosphorus, and selenium.
  • cycloheteroalkyl including heterocyclyl and heterocycle, includes a chain of atoms that includes both carbon and at least one heteroatom, such as heteroalkyl, and is optionally branched, where at least a portion of the chain is cyclic.
  • Illustrative heteroatoms include nitrogen, oxygen, and sulfur. In certain variations, illustrative heteroatoms also include phosphorus, and selenium.
  • Illustrative cycloheteroalkyl include, but are not limited to, tetrahydrofuryl, pyrrolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, homopiperazinyl, quinuclidinyl, and the like.
  • aryl includes monocyclic and polycyclic aromatic groups, including aromatic carbocyclic and aromatic heterocyclic groups, each of which may be optionally substituted.
  • carbocyclic and aromatic heterocyclic groups each of which may be optionally substituted.
  • carbocyclic groups each of which may be optionally substituted.
  • Illustrative aromatic carbocyclic groups described herein include, but are not limited to, phenyl, naphthyl, and the like.
  • heteroaryl includes aromatic heterocyclic groups, each of which may be optionally substituted.
  • Illustrative aromatic heterocyclic groups include, but are not limited to, pyridinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazinyl, quinolinyl, quinazolinyl, quinoxalinyl, thienyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, benzimidazolyl, benzoxazolyl, benzthiazolyl, benzisoxazolyl, benzisothiazolyl, and the like.
  • amino includes the group NH 2 , alkylamino, and dialkylamino, where the two alkyl groups in dialkylamino may be the same or different, i.e. alkylalkylamino.
  • amino includes methylamino, ethylamino, dimethylamino, methylethylamino, and the like.
  • amino modifies or is modified by another term, such as aminoalkyl, or acylamino the above variations of the term amino are included therein.
  • aminoalkyl includes H 2 N-alkyl, methylaminoalkyl, ethylaminoalkyl, dimethylaminoalkyl, methylethylaminoalkyl, and the like.
  • acylamino includes acylmethylamino, acylethylamino, and the like.
  • amino and derivatives thereof includes amino as described herein, and alkylamino, alkenylamino, alkynylamino, heteroalkylamino, heteroalkenylamino, heteroalkynylamino, cycloalkylamino, cycloalkenylamino, cycloheteroalkylamino, cycloheteroalkenylamino, arylamino, arylalkylamino, arylalkenylamino, arylalkynylamino, acylamino, and the like, each of which is optionally substituted.
  • amino derivative also includes urea, carbamate, and the like.
  • hydroxy and derivatives thereof includes OH, and alkyloxy, alkenyloxy, alkynyloxy, heteroalkyloxy, heteroalkenyloxy, heteroalkynyloxy, cycloalkyloxy, cycloalkenyloxy, cycloheteroalkyloxy, cycloheteroalkenyloxy, aryloxy, arylalkyloxy, arylalkenyloxy, arylalkynyloxy, acyloxy, and the like, each of which is optionally substituted.
  • hydroxy derivative also includes carbamate, and the like.
  • thio and derivatives thereof includes SH, and alkylthio, alkenylthio, alkynylthio, heteroalkylthio, heteroalkenylthio, heteroalkynylthio, cycloalkylthio, cycloalkenylthio, cycloheteroalkylthio, cycloheteroalkenylthio, arylthio, arylalkylthio, arylalkenylthio, arylalkynylthio, acylthio, and the like, each of which is optionally substituted.
  • thio derivative also includes thiocarbamate, and the like.
  • acyl includes formyl, and alkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, heteroalkylcarbonyl, heteroalkenylcarbonyl, heteroalkynylcarbonyl, cycloalkylcarbonyl, cycloalkenylcarbonyl, cycloheteroalkylcarbonyl, cycloheteroalkenylcarbonyl, arylcarbonyl, arylalkylcarbonyl, arylalkenylcarbonyl, arylalkynylcarbonyl, acylcarbonyl, and the like, each of which is optionally substituted.
  • carboxylate and derivatives thereof includes the group CO 2 H and salts thereof, and esters and amides thereof, and CN.
  • optionally substituted includes the replacement of hydrogen atoms with other functional groups on the radical that is optionally substituted.
  • Such other functional groups illustratively include, but are not limited to, amino, hydroxyl, halo, thiol, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof, and the like.
  • any of amino, hydroxyl, thiol, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, and/or sulfonic acid is optionally substituted.
  • the term “optionally substituted aryl” includes the replacement of hydrogen atoms with other functional groups on the aryl that is optionally substituted.
  • Such other functional groups illustratively include, but are not limited to, amino, hydroxyl, halo, thiol, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, nitro, sulfonic acids and derivatives thereof, carboxylic acids and derivatives thereof, and the like.
  • any of amino, hydroxyl, thiol, alkyl, haloalkyl, heteroalkyl, aryl, arylalkyl, arylheteroalkyl, and/or sulfonic acid is optionally substituted.
  • Illustrative substituents include, but are not limited to, a radical —(CH 2 ) x Z X , where x is an integer from 0-6 and Z X is selected from halogen, hydroxy, alkanoyloxy, including C 1 -C 6 alkanoyloxy, optionally substituted aroyloxy, alkyl, including C 1 -C 6 alkyl, alkoxy, including C 1 -C 6 alkoxy, cycloalkyl, including C 3 -C 8 cycloalkyl, cycloalkoxy, including C 3 -C 8 cycloalkoxy, alkenyl, including C 2 -C 6 alkenyl, alkynyl, including C 2 -C 6 alkynyl, haloalkyl, including C 1 -C 6 haloalkyl, haloalkoxy, including C 1 -C 6 haloalkoxy, halocycloalkyl, including C 3 -C 8
  • protecting group generally refers to chemical functional groups that can be selectively appended to and removed from functionality, such as amine groups, present in a chemical compound to render such functionality inert to chemical reaction conditions to which the compound is exposed. See. e.g., Greene and Wuts, Protective Groups in Organic Synthesis, 2d edition, John Wiley & Sons, New York, 1991. Numerous amine protecting groups are known in the art. Illustrative examples include the benzyloxycarbonyl (CBz), chlorobenzyloxycarbonyl, t-butyloxycarbonyl (Boc), fluorenylmethoxycarbonyl (Fmoc), isonicotinyloxycarbonyl (i-Noc) groups.
  • prodrug generally refers to any compound that when administered to a biological system generates a biologically active compound as a result of one or more spontaneous chemical reaction(s), enzyme-catalyzed chemical reaction(s), and/or metabolic chemical reaction(s), or a combination thereof.
  • the prodrug is typically acted upon by an enzyme (such as esterases, amidases, phosphatases, and the like), simple biological chemistry, or other process in vivo to liberate or regenerate the more pharmacologically active drug. This activation may occur through the action of an endogenous host enzyme or a non-endogenous enzyme that is administered to the host preceding, following, or during administration of the prodrug.
  • prodrug use is described in U.S. Pat. No. 5,627,165; and Pathalk et al., Enzymic protecting group techniques in organic synthesis, Stereosel. Biocatal. 775-797 (2000). It is appreciated that the prodrug is advantageously converted to the original drug as soon as the goal, such as targeted delivery, safety, stability, and the like is achieved, followed by the subsequent rapid elimination of the released remains of the group forming the prodrug.
  • Prodrugs may be prepared from the compounds described herein by attaching groups that ultimately cleave in vivo to one or more functional groups present on the compound, such as —OH—, —SH, —CO 2 H, —NR 2 .
  • Illustrative prodrugs include but are not limited to carboxylate esters where the group is alkyl, aryl, aralkyl, acyloxyalkyl, alkoxycarbonyloxyalkyl as well as esters of hydroxyl, thiol and amines where the group attached is an acyl group, an alkoxycarbonyl, aminocarbonyl, phosphate or sulfate.
  • esters also referred to as active esters, include but are not limited to 1-indanyl, N-oxysuccinimide; acyloxyalkyl groups such as acetoxymethyl, pivaloyloxymethyl, ⁇ -acetoxyethyl, ⁇ -pivaloyloxyethyl, 1-(cyclohexylcarbonyloxy)prop-1-yl, (1-aminoethyl)carbonyloxymethyl, and the like; alkoxycarbonyloxyalkyl groups, such as ethoxycarbonyloxymethyl, ⁇ -ethoxycarbonyloxyethyl, ⁇ -ethoxycarbonyloxyethyl, and the like; dialkylaminoalkyl groups, including di-lower alkylamino alkyl groups, such as dimethylaminomethyl, dimethylaminoethyl, diethylaminomethyl, diethylaminoethyl, and the like; 2-(alk
  • Further illustrative prodrugs contain a chemical moiety, such as an amide or phosphorus group functioning to increase solubility and/or stability of the compounds described herein.
  • Further illustrative prodrugs for amino groups include, but are not limited to, (C 3 -C 20 )alkanoyl; halo-(C 3 -C 20 )alkanoyl; (C 3 -C 20 )alkenoyl; (C 4 -C 7 )cycloalkanoyl; (C 3 -C 6 )-cycloalkyl(C 2 -C 16 )alkanoyl; optionally substituted aroyl, such as unsubstituted aroyl or aroyl substituted by 1 to 3 substituents selected from the group consisting of halogen, cyano, trifluoromethanesulphonyloxy, (C 1 -C 3 )alkyl and (C 1 -C 3 )alkoxy, each of which is
  • prodrugs themselves may not possess significant biological activity, but instead undergo one or more spontaneous chemical reaction(s), enzyme-catalyzed chemical reaction(s), and/or metabolic chemical reaction(s), or a combination thereof after administration in vivo to produce the compound described herein that is biologically active or is a precursor of the biologically active compound.
  • the prodrug is biologically active.
  • prodrugs may often serves to improve drug efficacy or safety through improved oral bioavailability, pharmacodynamic half-life, and the like.
  • Prodrugs also refer to derivatives of the compounds described herein that include groups that simply mask undesirable drug properties or improve drug delivery.
  • one or more compounds described herein may exhibit an undesirable property that is advantageously blocked or minimized may become pharmacological, pharmaceutical, or pharmacokinetic barriers in clinical drug application, such as low oral drug absorption, lack of site specificity, chemical instability, toxicity, and poor patient acceptance (bad taste, odor, pain at injection site, and the like), and others. It is appreciated herein that a prodrug, or other strategy using reversible derivatives, can be useful in the optimization of the clinical application of a drug.
  • the term “treating”, “contacting” or “reacting” when referring to a chemical reaction means to add or mix two or more reagents under appropriate conditions to produce the indicated and/or the desired product. It should be appreciated that the reaction which produces the indicated and/or the desired product may not necessarily result directly from the combination of two reagents which were initially added, i.e., there may be one or more intermediates which are produced in the mixture which ultimately leads to the formation of the indicated and/or the desired product.
  • composition generally refers to any product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combinations of the specified ingredients in the specified amounts. It is to be understood that the compositions described herein may be prepared from isolated compounds described herein or from salts, solutions, hydrates, solvates, and other forms of the compounds described herein. It is also to be understood that the compositions may be prepared from various amorphous, non-amorphous, partially crystalline, crystalline, and/or other morphological forms of the compounds described herein. It is also to be understood that the compositions may be prepared from various hydrates and/or solvates of the compounds described herein.
  • compositions that recite compounds described herein are to be understood to include each of, or any combination of, the various morphological forms and/or solvate or hydrate forms of the compounds described herein.
  • compositions may include one or more carriers, diluents, and/or excipients.
  • the compounds described herein, or compositions containing them, may be formulated in a therapeutically effective amount in any conventional dosage forms appropriate for the methods described herein.
  • compositions containing them may be administered by a wide variety of conventional routes for the methods described herein, and in a wide variety of dosage formats, utilizing known procedures (see generally, Remington: The Science and Practice of Pharmacy, (21 st ed., 2005)).
  • reaction was filtered to remove the imidazole HCl salt, extracted with de-ionized water, and the aqueous phase was back washed with dichloromethane, the combined organic phase was washed with brine, dried over Na 2 SO 4 , filtered to remove the Na 2 SO 4 , concentrated under reduced pressure, co-evaporated with toluene and dried under high-vacuum overnight to give 6.4 g of crude product 4 (vs 5.9 g of theoretical yield).
  • TES protected dipeptide was dissolved in 38 mL THF (anhydrous, inhibitor-free) and cooled to ⁇ 45° C. and stirred for 15 minutes before adding KHMDS (0.5 M in toluene, 25.5 mL, 12.8 mmol, 1.1 equiv) drop-wise. After the addition of KHMDS was complete, the reaction mixture was stirred at ⁇ 45° C. for 15 minutes, and chloromethyl butyrate (1.8 mL, 1.2 equiv, 14 mmol) was added. The reaction mixture changed from light yellow to a bluish color.
  • Tripeptide 6 Alkylated dipeptide 5 (4.3 g, 7.0 mmol), N-methyl pipecolinate (MEP) (4.0 g, 28.0 mmol, 4 equiv) and pentafluorophenol (5.7 g, 30.8 mmol 4.4 equiv) were added to a flask. N-methylpyrrolidone (NMP, 86 mL) was added to the mixture. To the mixture was added diisopropylcarbodiimide (DIC, 4.77 mL, 30.8 mmol, 4.4 equiv) was added to the mixture. The mixture was stirred at room temperature for 1 h. Pd/C (10%, dry, 1.7 g) was added.
  • the flask was shaken under hydrogen (30-35 psi) for 5 hours.
  • the reaction mixture was analyzed by HPLC. The starting material was found to be less than 3%.
  • the mixture was filtered through diatomaceous earth.
  • the diatomaceous earth was extracted with 200 mL ethyl acetate.
  • the filtrate and the ethyl acetate extract were combined and transferred to a separatory funnel and washed with 1% NaHCO 3 /10% NaCl solution (200 mL ⁇ 4).
  • the organic layer was isolated and evaporated on a rotary evaporator under reduced pressure.
  • the crude product was dissolved in 40 mL of MeOH/H 2 O (3:1).
  • the crude product solution was loaded onto a Biotage C18 column (Flash 65i, 350 g, 450 mL, 65 ⁇ 200 mm) and eluted with buffer A [10 mM NH 4 OAc/ACN (1:0] and B (ACN, acetonitrile). The fractions were collected and organic solvent was removed by evaporating on a rotary evaporator. 100 mL of 10% NaCl solution and 100 mL of methyl tert-butyl ether (MTBE) were added to the flask and the mixture was transferred to a separatory funnel. The organic layer was isolated and dried over anhydrous Na 2 SO 4 , filtered and evaporated on a rotary evaporator to dryness. 2.5 g of tripeptide intermediate 6 was obtained (yield 50%).
  • buffer A 10 mM NH 4 OAc/ACN (1:0]
  • B ACN, acetonitrile
  • Tripeptide 8 (229 mg, 0.367 mmol) was dissolved in EtOAc (anhydrous), 134.9 mg (0.733 mmol, 2.0 equiv) of pentafluorophenol was added, followed by 970 mg (1.84 mmol, 5.0 equiv) of DCC on the resin. The resulting reaction mixture was stirred at room temperature for 16 hours. LC-MS analysis indicated>96% conversion. The reaction mixture was filtered and concentrated to dryness, dried under high vacuum for 5 minutes.
  • reaction was filtered to remove the imidazole HCl salt, extracted with de-ionized water, and the aqueous phase was back-washed with dichloromethane, the combined organic phase was washed with brine, dried over Na 2 SO 4 , filtered to remove the Na 2 SO 4 , concentrated under reduced pressure, co-evaporated with toluene and dried under high-vacuum overnight to give 12.2 g of product 4.
  • TES protected dipeptide was dissolved in 80 mL THF (anhydrous, inhibitor-free) and cooled to ⁇ 45° C. and stirred for 15 minutes before adding KHMDS (0.5 M in toluene, 50 mL, 25.0 mmol, 1.05 equiv) drop-wise. After the addition of KHMDS was complete, the reaction mixture was stirred at ⁇ 45° C. for 15 minutes, and chloromethyl butyrate (3.6 mL, 1.2 equiv, 28.3 mmol) was added. The reaction mixture changed from light yellow to a bluish color.
  • Tripeptide 6 Alkylated dipeptide 5 (7.6 g, 12.4 mmol), N-methyl pipecolinate (MEP) (7.0 g, 48.9 mmol, 4 equiv) and pentafluorophenol (10.0 g, 54.3 mmol 4.4 equiv) were added to a flask. N-methylpyrrolidone (NMP, 152 mL) was added to the mixture. To the mixture was added diisopropylcarbodiimide (DIC, 8.43 mL, 54.4 mmol, 4.4 equiv) was added to the mixture. The mixture was stirred at room temperature for 1 h. Pd/C (10%, dry, 3.0 g) was added.
  • the flask was shaken under hydrogen (30-35 psi) for 5 hours.
  • the reaction mixture was analyzed by HPLC.
  • the reaction was complete.
  • the mixture was filtered through celite.
  • the celite was washed with 500 mL ethyl acetate.
  • the solutions were combined and transferred to a separatory funnel and washed with 1% NaHCO 3 /10% NaCl solution (250 mL ⁇ 4).
  • the organic layer was isolated and evaporated on a rotary evaporator under reduced pressure.
  • the crude product was dissolved in dichloromethane and the urea was filtered.
  • reaction mixture was drained from the reactor in 1 L portions and extracted with 94% EtOAc-6% MeOH (vol./vol.) solution (3 ⁇ 1 L). The combined extracts were washed with brine, dried over Na 2 SO 4 and concentrated under reduced pressure. 2.8 g of product 6 was recovered (95%). The product appeared to be clean by UPLC analysis, except for pentafluorophenol carried over from the previous reaction.
  • TES protected dipeptide was dissolved in 100 mL THF (anhydrous, inhibitor-free), cooled to ⁇ 45° C., and stirred at ⁇ 45° C. for 15 minutes before adding KHMDS (0.5 M in toluene, 61 mL, 1.05 equiv.), drop-wise. After the addition of KHMDS was finished, the reaction was stirred at ⁇ 45° C. for 20 minutes, and chloromethyl butyrate (4.4 mL, 1.1 equiv.) was added. The reaction mixture was stirred at ⁇ 45° C. for another 20 minutes. The reaction was quenched with 25 mL MeOH and warmed to room temperature.
  • KHMDS 0.5 M in toluene, 61 mL, 1.05 equiv.
  • TFA was removed under reduced pressure, co-evaporated with CH 2 Cl 2 (3 ⁇ 30 mL) and activated Tut-derivative 9 was dried under high vacuum for 18 h.
  • the tripeptide acid 8 (2.51 g, 4.02 mmol) was dissolved in 70 mL CH 2 Cl 2 (anhydrous) and 1.48 g (8.04 mmol, 2.0 equiv) of pentafluorophenol in 5 mL of CH 2 Cl 2 was added, followed by 8.74 g (20.1 mmol, 5.0 equiv) of DCC-resin. The resulting reaction mixture was stirred at room temperature for 20 hours.
  • a binding ligand-linker intermediate containing a thiol group is taken in deionized water (ca. 20 mg/mL, bubbled with argon for 10 minutes prior to use) and the pH of the suspension was adjusted with aqueous phosphate (bubbled with argon for 10 minutes prior to use) to a pH of about 7.0 (the suspension may become a solution when the pH increased). Additional deionized water is added (ca. 20-25%) to the solution as needed, and to the aqueous solution is added immediately a solution of compound (2) in acetonitrile (ca. 20 mg/mL). The reaction mixture becomes homogenous quickly.
  • the reaction mixture is diluted with 2.0 mM sodium phosphate buffer (pH 7.0, ca 150 volume percent) and the acetonitrile is removed under vacuum. The resulting suspension is filtered and the filtrate may be purified by preparative HPLC. Fractions are lyophilized to isolate the conjugates.
  • the foregoing method is equally applicable for preparing other tubulysin conjugates by the appropriate selection of the tubulysin starting compound, including tubulysin starting compounds having a 3-nitropyridin-2-ylthio activating group.
  • This binding ligand-linker intermediate was prepared by SPPS according to the general peptide synthesis procedure described herein starting from H-Cys(4-methoxytrityl)-2-chlorotrityl-Resin, and the following SPPS reagents:
  • Coupling steps In a peptide synthesis vessel add the resin, add the amino acid solution, DIPEA, and PyBOP. Bubble argon for 1 hr. and wash 3 ⁇ with DMF and IPA. Use 20% piperidine in DMF for Fmoc deprotection, 3 ⁇ (10 min), before each amino acid coupling. Continue to complete all 9 coupling steps. At the end treat the resin with 2% hydrazine in DMF 3 ⁇ (5 min) to cleave TFA protecting group on Pteroic acid, wash the resin with DMF (3 ⁇ ), IPA (3 ⁇ ), MeOH (3 ⁇ ), and bubble the resin with argon for 30 min.
  • Cleavage step Reagent: 92.5% TFA, 2.5% H2O, 2.5% triisopropylsilane, 2.5% ethanedithiol.
  • EC0531 is prepared according to the processes described herein from compound (2) and EC0488 in 73% yield.
  • the ether analog of compound 103 can also be prepared. Reductive condensation of that ether analog with MEP yields 105 directly.
  • reaction was filtered to remove the imidazole HCl salt, extracted with de-ionized water, and the aqueous phase was back-washed with dichloromethane. The combined organic phase was washed with brine, dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure. The resulting residue was co-evaporated with toluene and dried over high-vacuum overnight to give 6.4 g of crude product 102 (5.9 g theoretical yield).
  • the flask was shaken under hydrogen (30-35 psi) for 5 hours.
  • the reaction mixture was analyzed by HPLC. The starting material was found to be less than 3%.
  • the mixture was filtered through celite. The celite was washed with ethyl acetate (200 mL). The combined filtrate was transferred to separatory funnel and washed with 1% NaHCO 3 /10% NaCl solution (200 mL ⁇ 4). The organic layer was isolated and rotavaped under reduced pressure.
  • the crude product was dissolved in 40 mL of MeOH/H 2 O (3:1).
  • the crude product solution was loaded onto a Biotage C18 column (Flash 65i, 350 g, 450 mL, 65 ⁇ 200 mm) and eluted with buffer A [10 mM NH 4 OAc/ACN (1:1)] and B (ACN). The fractions were collected and organic solvent was removed under reduced pressure. A 10% NaCl solution (100 mL) and methyl t-butyl ether (MTBE) (100 mL) were added to the flask and the mixture was transferred to a separatory funnel. The organic layer was isolated and dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure. 2.5 g of tripeptide intermediate 104 was obtained (yield 50%).
  • reaction mixture was filtered and concentrated under reduced pressure, and the residue was dissolved in a solution of N,N-dimethylformamide (2 mL) and N,N-diisopropylethylamine (8mL, 0.046 mmol).
  • PFP ester intermediate (6.0 mg, 0.023 mmol) was added and the reaction mixture was stirred at room temperature for 2 h under argon.
  • LCMS (10-100% ACN, 50 mM NH 4 HCO 3 pH7) indicated all of the activated intermediate had been consumed and product had been formed.
  • the reaction mixture was purified by preparative HPLC (10-100% ACN, 50 mM NH 4 HCO 3 pH7) to yield 108b (4.7 mg, 37%).
  • reaction mixture was filtered and concentrated under reduced pressure, and the residue was dissolved in a solution of N,N-dimethylformamide (2 mL) and N,N-diisopropylethylamine (4 ⁇ L, 0.023 mmol).
  • PFP ester intermediate (3.0 mg, 0.012 mmol) was added and the reaction mixture was stirred at room temperature for 2 h under Argon.
  • LC-MS (10-100% ACN, 50 mM NH 4 HCO 3 pH7) indicated all of the activated intermediate had been consumed and product had been formed.
  • the reaction mixture was purified by preparative HPLC (10-100% ACN, 50 mM NH 4 HCO 3 pH7) to yield 108c (1.1 mg, 17%).
  • Step 2 EC1426 (114 mg) was dissolved in anhydrous DCM (1.5 mL) and to which was added TFA (0.50 mL). The resulting solution was stirred at ambient temperature under argon for 70 minutes and concentrated under reduced pressure to give a residue, which was co-evaporated with anhydrous DCM (2.0 mL ⁇ 3) and vacuumed at ambient temperature for 9 hours prior to use in Step 3.
  • Step 3 The residue from Step 1 was dissolved in anhydrous DCM (1.5 mL) and to this solution was added DIPEA (0.50 mL) followed by a solution of the residue from Step 2 dissolved in anhydrous dimethylformamide (DMF, 1.5 mL). The resulting solution was stirred at ambient temperature under argon for 1 hour, diluted with ethyl acetate (EtOAc, 60 mL), and washed with brine (20 mL ⁇ 3).
  • EtOAc ethyl acetate
  • the ether analogs of compounds III and 112 can be used to prepare the tubulysin intermediates described herein.

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9505747B2 (en) 2012-03-29 2016-11-29 Endocyte, Inc. Processes for preparing tubulysin derivatives and conjugates thereof
US9550734B2 (en) 2004-07-23 2017-01-24 Endocyte, Inc. Bivalent linkers and conjugates thereof
US9555139B2 (en) 2007-03-14 2017-01-31 Endocyte, Inc. Binding ligand linked drug delivery conjugates of tubulysins
WO2017031209A1 (en) * 2015-08-18 2017-02-23 Endocyte, Inc. Tubulysin analogs and methods
US9662402B2 (en) 2012-10-16 2017-05-30 Endocyte, Inc. Drug delivery conjugates containing unnatural amino acids and methods for using
US9877965B2 (en) 2007-06-25 2018-01-30 Endocyte, Inc. Vitamin receptor drug delivery conjugates for treating inflammation
US10080805B2 (en) 2012-02-24 2018-09-25 Purdue Research Foundation Cholecystokinin B receptor targeting for imaging and therapy
US10738086B2 (en) 2007-06-25 2020-08-11 Endocyte Inc. Conjugates containing hydrophilic spacer linkers
US11229708B2 (en) 2015-12-04 2022-01-25 Seagen Inc. Conjugates of quaternized tubulysin compounds
US11793880B2 (en) 2015-12-04 2023-10-24 Seagen Inc. Conjugates of quaternized tubulysin compounds

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
BR112015011118B1 (pt) 2012-11-15 2022-12-13 Endocyte, Inc Conjugado; composição farmacêutica; e uso de um conjugado
US20140249315A1 (en) * 2013-03-01 2014-09-04 Endocyte, Inc. Processes for preparing tubulysins
WO2015113760A1 (en) * 2014-01-28 2015-08-06 Tube Pharmaceuticals Gmbh Cytotoxic tubulysin compounds for conjugation
EP3131587A4 (en) * 2014-04-14 2017-11-01 Endocyte, Inc. Drug delivery conjugates for treating resistant cancer and for use in combination therapy
US20160074527A1 (en) * 2014-09-17 2016-03-17 Genentech, Inc. Pyrrolobenzodiazepines and antibody disulfide conjugates thereof
HK1248069A1 (zh) * 2015-02-25 2018-10-12 William Marsh Rice University 去乙酰氧基微管溶素h及其类似物
US11274124B2 (en) 2017-11-29 2022-03-15 William Marsh Rice University Tubulysin analogues as anticancer agents and payloads for antibody-drug conjugates and methods of treatment therewith
MA53765B2 (fr) * 2018-12-21 2025-04-30 Regeneron Pharmaceuticals, Inc. Tubulysines et conjugués tubulysines-protéines

Family Cites Families (205)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2515483A (en) 1946-08-10 1950-07-18 Merck & Co Inc Diacylated pteroic acid and process for preparing same
US2816110A (en) 1956-11-23 1957-12-10 Merck & Co Inc Methods for the production of substituted pteridines
US3392173A (en) 1964-03-09 1968-07-09 Lilly Co Eli Novel acyl derivatives of desacetyl-vincaleukoblastine and processes for their preparation
US3387001A (en) 1964-10-19 1968-06-04 Lilly Co Eli Novel aminoacyl esters of desacetyl vincaleukoblastine
US4203898A (en) 1977-08-29 1980-05-20 Eli Lilly And Company Amide derivatives of VLB, leurosidine, leurocristine and related dimeric alkaloids
US4166810A (en) 1978-04-20 1979-09-04 Eli Lilly And Company Derivatives of 4-desacetyl VLB C-3 carboxyhydrazide
US4337339A (en) 1979-04-30 1982-06-29 Baker Instruments Corp. Process for preparation of folic acid derivatives
US4639456A (en) 1980-06-10 1987-01-27 Omnichem S.A. Vinblastin-23-oyl amino acid derivatives
US4316885A (en) 1980-08-25 1982-02-23 Ayerst, Mckenna And Harrison, Inc. Acyl derivatives of rapamycin
US4713249A (en) 1981-11-12 1987-12-15 Schroeder Ulf Crystallized carbohydrate matrix for biologically active substances, a process of preparing said matrix, and the use thereof
US5140104A (en) 1982-03-09 1992-08-18 Cytogen Corporation Amine derivatives of folic acid analogs
EP0116208B1 (en) 1982-12-07 1988-03-30 Kyowa Hakko Kogyo Co., Ltd. Mitomycin analogues
ATE64396T1 (de) 1983-04-29 1991-06-15 Omnichem Sa Konjugierte vinblastin-verbindungen und ihre derivate, verfahren zu ihrer herstellung und diese enthaltende pharmazeutische zusammensetzungen.
JPS59175493U (ja) 1983-05-12 1984-11-22 株式会社小松製作所 レ−ザ加工装置
US4866180A (en) 1984-02-24 1989-09-12 Bristol-Myers Company Amino disulfide thiol exchange products
GB8413849D0 (en) 1984-05-31 1984-07-04 Amersham Int Plc Nmr contrast agents
JPS60255789A (ja) 1984-06-01 1985-12-17 Kyowa Hakko Kogyo Co Ltd マイトマイシン誘導体，その製造法および抗腫瘍剤
US5266333A (en) 1985-03-06 1993-11-30 American Cyanamid Company Water dispersible and water soluble carbohydrate polymer compositions for parenteral administration of growth hormone
US4650803A (en) 1985-12-06 1987-03-17 University Of Kansas Prodrugs of rapamycin
IL82579A0 (en) 1986-05-27 1987-11-30 Lilly Co Eli Immunoglobulin conjugates
US4801688A (en) 1986-05-27 1989-01-31 Eli Lilly And Company Hydrazone immunoglobulin conjugates
US5103018A (en) 1986-08-26 1992-04-07 Kyowa Hakko Kogyo Kabushiki Kaisha Mitomycin derivatives
US5094849A (en) 1988-08-08 1992-03-10 Eli Lilly And Company Cytotoxic antibody conjugates of hydrazide derivatized vinca analogs via simple organic linkers
US5006652A (en) 1988-08-08 1991-04-09 Eli Lilly And Company Intermediates for antibody-vinca drug conjugates
US5688488A (en) 1989-04-03 1997-11-18 Purdue Research Foundation Composition and method for tumor imaging
US5108921A (en) 1989-04-03 1992-04-28 Purdue Research Foundation Method for enhanced transmembrane transport of exogenous molecules
CA2044590A1 (en) 1989-11-13 1991-05-14 Marc D. Better Chimeric mouse-human a10 antibody with specificity to a human tumor cell antigen
US5627165A (en) 1990-06-13 1997-05-06 Drug Innovation & Design, Inc. Phosphorous prodrugs and therapeutic delivery systems using same
US5998603A (en) 1994-09-29 1999-12-07 Isis Pharmaceuticals, Inc. 4'-desmethyl nucleoside analogs, and oligomers thereof
WO1992003569A1 (en) 1990-08-29 1992-03-05 Centre Hospitalier Regional De Nantes Protein polyligands joined to a stable protein core
US5221670A (en) 1990-09-19 1993-06-22 American Home Products Corporation Rapamycin esters
US5378696A (en) 1990-09-19 1995-01-03 American Home Products Corporation Rapamycin esters
US5130307A (en) 1990-09-28 1992-07-14 American Home Products Corporation Aminoesters of rapamycin
US5233036A (en) 1990-10-16 1993-08-03 American Home Products Corporation Rapamycin alkoxyesters
US5120842A (en) 1991-04-01 1992-06-09 American Home Products Corporation Silyl ethers of rapamycin
US5100883A (en) 1991-04-08 1992-03-31 American Home Products Corporation Fluorinated esters of rapamycin
US5194447A (en) 1992-02-18 1993-03-16 American Home Products Corporation Sulfonylcarbamates of rapamycin
US5118678A (en) 1991-04-17 1992-06-02 American Home Products Corporation Carbamates of rapamycin
US5138051A (en) 1991-08-07 1992-08-11 American Home Products Corporation Rapamycin analogs as immunosuppressants and antifungals
US5118677A (en) 1991-05-20 1992-06-02 American Home Products Corporation Amide esters of rapamycin
US5169851A (en) 1991-08-07 1992-12-08 American Home Products Corporation Rapamycin analog as immunosuppressants and antifungals
US6335434B1 (en) 1998-06-16 2002-01-01 Isis Pharmaceuticals, Inc., Nucleosidic and non-nucleosidic folate conjugates
US5151413A (en) 1991-11-06 1992-09-29 American Home Products Corporation Rapamycin acetals as immunosuppressant and antifungal agents
US5159079A (en) 1991-12-20 1992-10-27 Eli Lilly And Company 2-piperidones as intermediates for 5-deaza-10-oxo- and 5-deaza-10-thio-5,6,7,8-tetrahydrofolic acids
US6004555A (en) 1992-03-05 1999-12-21 Board Of Regents, The University Of Texas System Methods for the specific coagulation of vasculature
US6022966A (en) 1993-11-22 2000-02-08 Neorx Corporation Pretargeting methods and compounds
US5302584A (en) 1992-10-13 1994-04-12 American Home Products Corporation Carbamates of rapamycin
US5258389A (en) 1992-11-09 1993-11-02 Merck & Co., Inc. O-aryl, O-alkyl, O-alkenyl and O-alkynylrapamycin derivatives
US5260300A (en) 1992-11-19 1993-11-09 American Home Products Corporation Rapamycin carbonate esters as immuno-suppressant agents
US5583020A (en) 1992-11-24 1996-12-10 Ribozyme Pharmaceuticals, Inc. Permeability enhancers for negatively charged polynucleotides
US7279561B1 (en) 1993-04-23 2007-10-09 Wyeth Anti-rapamycin monoclonal antibodies
DE69435044T2 (de) 1993-04-23 2008-09-18 Wyeth Rapamycin - Konjugate und Antikörper
US5562907A (en) 1993-05-14 1996-10-08 Arnon; Stephen S. Method to prevent side-effects and insensitivity to the therapeutic uses of toxins
US5391730A (en) 1993-10-08 1995-02-21 American Home Products Corporation Phosphorylcarbamates of rapamycin and oxime derivatives thereof
US5385909A (en) 1993-11-22 1995-01-31 American Home Products Corporation Heterocyclic esters of rapamycin
US5385908A (en) 1993-11-22 1995-01-31 American Home Products Corporation Hindered esters of rapamycin
US5385910A (en) 1993-11-22 1995-01-31 American Home Products Corporation Gem-distributed esters of rapamycin
US5389639A (en) 1993-12-29 1995-02-14 American Home Products Company Amino alkanoic esters of rapamycin
US5417982A (en) 1994-02-17 1995-05-23 Modi; Pankaj Controlled release of drugs or hormones in biodegradable polymer microspheres
IL112873A (en) 1994-03-08 2005-03-20 Wyeth Corp Rapamycin-fkbp12 binding proteins, their isolation and their use
US6171859B1 (en) 1994-03-30 2001-01-09 Mitokor Method of targeting conjugate molecules to mitochondria
US5362718A (en) 1994-04-18 1994-11-08 American Home Products Corporation Rapamycin hydroxyesters
US5463048A (en) 1994-06-14 1995-10-31 American Home Products Corporation Rapamycin amidino carbamates
US5491231A (en) 1994-11-28 1996-02-13 American Home Products Corporation Hindered N-oxide esters of rapamycin
US5547668A (en) 1995-05-05 1996-08-20 The Board Of Trustees Of The University Of Illinois Conjugates of folate anti-effector cell antibodies
AUPN449295A0 (en) 1995-07-28 1995-08-24 Inner And Eastern Health Care Network, The Radioprotectors
US6207157B1 (en) 1996-04-23 2001-03-27 The United States Of America As Represented By The Department Of Health And Human Services Conjugate vaccine for nontypeable Haemophilus influenzae
US6030941A (en) 1996-05-01 2000-02-29 Avi Biopharma, Inc. Polymer composition for delivering substances in living organisms
AU717020B2 (en) 1996-05-03 2000-03-16 Immunomedics Inc. Targeted combination immunotherapy of cancer
DE19621133A1 (de) 1996-05-24 1997-11-27 Boehringer Mannheim Gmbh Bestimmungsverfahren mit oligomerisierten Rezeptoren
ATE298344T1 (de) 1996-08-27 2005-07-15 Univ Utah Res Found Biokonjugate und verabreichung von bioaktiven substanzen
WO1998010651A1 (en) 1996-09-12 1998-03-19 Merck & Co., Inc. Conjugates useful in the treatment of prostate cancer
US6056973A (en) 1996-10-11 2000-05-02 Sequus Pharmaceuticals, Inc. Therapeutic liposome composition and method of preparation
US6177404B1 (en) 1996-10-15 2001-01-23 Merck & Co., Inc. Conjugates useful in the treatment of benign prostatic hyperplasia
US6071532A (en) 1996-10-15 2000-06-06 Emory University Synthesis of glycophospholipid and peptide-phospholipid conjugates and uses thereof
US6291673B1 (en) 1997-10-17 2001-09-18 Purdue Research Foundation Folic acid derivatives
GB9723669D0 (en) 1997-11-07 1998-01-07 Univ Aberdeen Skin penetration enhancing components
US6399638B1 (en) 1998-04-21 2002-06-04 Bristol-Myers Squibb Company 12,13-modified epothilone derivatives
US6093382A (en) 1998-05-16 2000-07-25 Bracco Research Usa Inc. Metal complexes derivatized with folate for use in diagnostic and therapeutic applications
ES2347027T3 (es) 1998-05-22 2010-10-22 The Board Of Trustees Of The Leland Stanford Junior University Moleculas bifuncionales y terapias basadas en las mismas.
US6974581B1 (en) 1998-12-15 2005-12-13 Aventis Pasteur Limited Multi-component vaccine comprising at least two antigens from haemophilus influenzae to protect against disease
US6291684B1 (en) 1999-03-29 2001-09-18 Bristol-Myers Squibb Company Process for the preparation of aziridinyl epothilones from oxiranyl epothilones
KR100669053B1 (ko) 1999-04-23 2007-01-15 알자 코포레이션 리포좀에 사용하기 위한 절단가능 결합을 갖는 콘쥬게이트
AUPQ014799A0 (en) 1999-05-04 1999-05-27 Access Pharmaceuticals Australia Pty Limited Amplification of folate-mediated targeting to tumor cells using polymers
AUPQ071299A0 (en) 1999-06-02 1999-06-24 Access Pharmaceuticals Australia Pty Limited Vitamin directed dual targeting therapy
US6822086B1 (en) 1999-08-09 2004-11-23 The General Hospital Corporation Drug-carrier complexes and methods of use thereof
CA2381425A1 (en) 1999-08-24 2001-03-01 Cellgate, Inc. Enhancing drug delivery across and into epithelial tissues using oligo arginine moieties
JP2003512338A (ja) 1999-10-15 2003-04-02 メイオウ・フアウンデーシヨン・フオー・メデイカル・エジユケイシヨン・アンド・リサーチ 造影剤および抗腫瘍薬として有用なコバラミン結合体
US7067111B1 (en) 1999-10-25 2006-06-27 Board Of Regents, University Of Texas System Ethylenedicysteine (EC)-drug conjugates, compositions and methods for tissue specific disease imaging
ES2276708T3 (es) 1999-11-24 2007-07-01 Immunogen, Inc. Agentes citotoxicos que comprenden taxanos y su uso terapeutico.
PL211872B1 (pl) 2000-03-31 2012-07-31 Purdue Research Foundation Kompozycja farmaceutyczna
GB0008090D0 (en) 2000-04-04 2000-05-24 Morgan Samuels Limited Trouser waist adjuster
US6670355B2 (en) 2000-06-16 2003-12-30 Wyeth Method of treating cardiovascular disease
US6290929B1 (en) 2000-07-28 2001-09-18 The Procter & Gamble Company Cancer treatment
JP2004507465A (ja) 2000-08-11 2004-03-11 ワイス エストロゲン受容体陽性癌腫の治療方法
EP1319008B1 (en) 2000-09-19 2008-10-15 Wyeth Water soluble rapamycin esters
US6399625B1 (en) 2000-09-27 2002-06-04 Wyeth 1-oxorapamycins
US6399626B1 (en) 2000-10-02 2002-06-04 Wyeth Hydroxyesters of 7-desmethylrapamycin
US6440991B1 (en) 2000-10-02 2002-08-27 Wyeth Ethers of 7-desmethlrapamycin
IL155952A0 (en) 2000-11-28 2003-12-23 Wyeth Corp Expression analysis of fkbp nucleic acids and polypeptides useful in the diagnosis and treatment of prostate cancer
US20020168737A1 (en) 2001-01-24 2002-11-14 Cornish Virginia W. Binding and catalysis screen for high throughput determination of protein function using chemical inducers of dimerization
EP2388590A1 (en) 2001-04-02 2011-11-23 Dana Farber Cancer Institute PD-1, a receptor for B7-4, and uses thereof
AR036993A1 (es) 2001-04-02 2004-10-20 Wyeth Corp Uso de agentes que modulan la interaccion entre pd-1 y sus ligandos en la submodulacion de respuestas inmunologicas
EP1389209B1 (en) 2001-04-24 2009-04-08 Purdue Research Foundation Folate mimetics and folate-receptor binding conjugates thereof
CA2445826C (en) 2001-05-02 2014-03-25 Purdue Research Foundation Treatment and diagnosis of macrophage mediated disease
US7109165B2 (en) 2001-05-18 2006-09-19 Sirna Therapeutics, Inc. Conjugates and compositions for cellular delivery
MXPA03010909A (es) 2001-06-01 2004-02-17 Bristol Myers Squibb Co Derivados de epotilona.
US20040018203A1 (en) 2001-06-08 2004-01-29 Ira Pastan Pegylation of linkers improves antitumor activity and reduces toxicity of immunoconjugates
UA77200C2 (en) 2001-08-07 2006-11-15 Wyeth Corp Antineoplastic combination of cci-779 and bkb-569
MXPA04001523A (es) 2001-08-22 2004-05-31 Wyeth Corp Dialdehidos de rapamicina.
BR0211986A (pt) 2001-08-22 2004-09-28 Wyeth Corp 29-enóis de rapamicina
KR20040053136A (ko) 2001-09-28 2004-06-23 펄듀 리서치 파운데이션 리간드-면역원 공액체를 이용한 치료 방법
GR1004163B (el) 2001-11-01 2003-02-21 Πολυκυκλικα παραγωγα τροποποιησης των οπτικων ιδιοτητων και των ιδιοτητων αντοχης στο πλασμα των πολυμερων λιθογραφιας
US20050074457A1 (en) 2001-12-12 2005-04-07 Adeela Kamal Assays and implements for determining and modulating hsp90 binding activity
US20030194409A1 (en) 2002-01-17 2003-10-16 Rothman James E. Conjugate heat shock protein-binding peptides
US8043602B2 (en) 2002-02-07 2011-10-25 Endocyte, Inc. Folate targeted enhanced tumor and folate receptor positive tissue optical imaging technology
US7000695B2 (en) 2002-05-02 2006-02-21 Halliburton Energy Services, Inc. Expanding wellbore junction
DK2260875T3 (da) 2002-05-06 2014-06-30 Endocyte Inc Folatreceptor-targetede billeddannelsesmidler
US6596757B1 (en) 2002-05-14 2003-07-22 Immunogen Inc. Cytotoxic agents comprising polyethylene glycol-containing taxanes and their therapeutic use
WO2003097647A1 (en) 2002-05-15 2003-11-27 Endocyte, Inc. Vitamin-mitomycin conjugates
US7776814B2 (en) 2002-07-09 2010-08-17 R&D-Biopharmaceuticals Gmbh Tubulysin conjugates
AU2003266233A1 (en) 2002-07-09 2004-01-23 Morphochem Aktiengesellschaft Fur Kombinatorische Chemie Novel tubulysin analogues
HRP20050186A2 (en) 2002-07-31 2005-10-31 Schering Aktiengesellschaft New effector conjugates, process for their production and their pharmaceutical use
DE10241152A1 (de) 2002-09-05 2004-03-18 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Tubulysin-Biosynthese-Gene
BRPI0314042B8 (pt) 2002-09-06 2021-05-25 Calando Pharmaceuticals Inc polímeros à base de ciclodextrina para o fornecimento de agentes terapêuticos ligados a eles por covalência
US20040047917A1 (en) 2002-09-06 2004-03-11 Stephen Wilson Drug delivery and targeting with vitamin B12 conjugates
US7122361B2 (en) 2002-10-10 2006-10-17 Wyeth Compositions employing a novel human kinase
EP1558137A2 (en) 2002-10-24 2005-08-03 Research Corporation Technologies Functional mri agents for cancer imaging
DE10254439A1 (de) 2002-11-21 2004-06-03 GESELLSCHAFT FüR BIOTECHNOLOGISCHE FORSCHUNG MBH (GBF) Tubulysine, Herstellungsverfahren und Tubulysin-Mittel
EP1578197A4 (en) 2002-11-21 2006-05-17 Wyeth Corp COMPOSITION AND METHOD FOR THE TREATMENT OF LUPUS NEPHRITIS
EP1572242B1 (en) 2002-12-13 2014-04-16 Immunomedics, Inc. Immunoconjugates with an intracellularly-cleavable linkage
DK1592457T3 (da) 2003-01-27 2012-10-22 Endocyte Inc Folat-vinblastin-konjugat som lægemiddel
AR042938A1 (es) 2003-02-06 2005-07-06 Wyeth Corp Uso del cci-779 en el tratamiento de la fibrosis hepatica
DK1620544T3 (en) 2003-04-17 2019-01-14 Alnylam Pharmaceuticals Inc MODIFIED iRNA AGENTS
WO2004101803A2 (en) 2003-05-12 2004-11-25 Wyeth Holdings Corporation Process for producing anticancer agent ll-d45042
WO2005010010A1 (en) 2003-07-16 2005-02-03 Wyeth Cci-779 isomer c
KR20060057605A (ko) 2003-08-07 2006-05-26 와이어쓰 Ｃｃｉ-779의 위치선택적 합성 방법
DE602005003453T2 (de) 2004-01-30 2008-09-25 Bayer Schering Pharma Aktiengesellschaft Neue effektor-konjugate, verfahren zu ihrer herstellung und ihre pharmazeutische verwendung
US20060105975A1 (en) 2004-04-19 2006-05-18 Shannon Pendergrast Aptamer-mediated intracellular delivery of therapeutic oligonucleotides
JP4806680B2 (ja) 2004-05-19 2011-11-02 メダレックス インコーポレイテッド 自己犠牲リンカー及び薬剤複合体
WO2005115912A1 (ja) 2004-05-25 2005-12-08 Matsushita Electric Industrial Co., Ltd. 水素生成装置及びそれを用いた燃料電池システム
JP5149620B2 (ja) 2004-07-23 2013-02-20 エンドサイト，インコーポレイテッド ２価リンカーおよびその結合体
EP1791567B1 (en) 2004-08-10 2015-07-29 Alnylam Pharmaceuticals Inc. Chemically modified oligonucleotides
AU2005294214A1 (en) 2004-10-07 2006-04-20 Emory University Multifunctional nanoparticles conjugates and their use
US20060222653A1 (en) 2004-11-12 2006-10-05 Xencor, Inc. Antibodies operably linked to selected chemoattractants
US20110166319A1 (en) 2005-02-11 2011-07-07 Immunogen, Inc. Process for preparing purified drug conjugates
TW200640493A (en) 2005-02-16 2006-12-01 Insert Therapeutics Inc Cyclodextrin-based polymers for therapeutics delivery
US7312217B2 (en) 2005-03-11 2007-12-25 Syntrix Biosystems, Inc. Aminopterin dosage forms and methods for inflammatory disorders
EP1863816B1 (en) 2005-03-16 2014-06-25 Endocyte, Inc. Synthesis and purification of pteroic acid and conjugates thereof
ATE460668T1 (de) 2005-03-30 2010-03-15 Purdue Research Foundation Verfahren zur prognose von brustkrebs mittels quantifizierung von zellulären folat vitamin rezeptoren
NZ565075A (en) 2005-06-20 2011-05-27 Psma Dev Company Llc PSMA antibody-drug conjugates
US20070009434A1 (en) 2005-07-05 2007-01-11 Low Philip S Imaging and therapeutic method using monocytes
EP1937719A4 (en) 2005-08-19 2010-11-24 Novo Nordisk As GLYCOPEGYLATED FACTOR VII AND FACTOR VIIA
EP2382995A3 (en) 2005-08-19 2013-09-25 Endocyte, Inc. Ligand conjugates of Vinca alkaloids, analogs and derivatives
AU2006279304A1 (en) 2005-08-19 2007-02-22 Endocyte, Inc. Multi-drug ligand conjugates
EP3539572A1 (en) 2005-08-24 2019-09-18 ImmunoGen, Inc. Process for preparing maytansinoid antibody conjugates
WO2008048298A2 (en) 2005-11-21 2008-04-24 Medivas, Llc Polymer particles for delivery of macromolecules and methods of use
PE20080102A1 (es) 2006-05-25 2008-02-11 Bristol Myers Squibb Co Conjugados de analogos de aziridinil-epotilona y composiciones farmaceuticas que comprenden los mismos
JP2010509570A (ja) 2006-11-03 2010-03-25 パーデュー・リサーチ・ファウンデーション エクスビボフローサイトメトリーの方法および装置
US20080176958A1 (en) 2007-01-24 2008-07-24 Insert Therapeutics, Inc. Cyclodextrin-based polymers for therapeutics delivery
WO2008101231A2 (en) 2007-02-16 2008-08-21 Endocyte, Inc. Methods and compositions for treating and diagnosing kidney disease
CN104127878A (zh) 2007-03-14 2014-11-05 恩多塞特公司 结合配体连接的微管溶素递药缀合物
US8980833B2 (en) 2007-05-10 2015-03-17 R&D-Biopharmaceuticals Gmbh Tubulysine derivatives
BRPI0812970A2 (pt) 2007-06-25 2019-09-24 Endocyte Inc conjugados contendo espaçadores hidrofílicos
US8476451B2 (en) 2007-07-20 2013-07-02 The Regents Of The University Of California Tubulysin D analogues
EP4464384A3 (en) 2007-08-17 2025-01-08 Purdue Research Foundation Psma binding ligand-linker conjugates and methods for using
US9187521B2 (en) 2007-10-25 2015-11-17 Endocyte, Inc. Tubulysins and processes for preparing
EP2231194B1 (en) 2007-12-04 2017-02-22 Alnylam Pharmaceuticals Inc. Folate-irna conjugates
EP2276506A4 (en) 2008-04-30 2014-05-07 Immunogen Inc EFFICIENT CONJUGATES AND HYDROPHILIC BINDER
US20100040669A1 (en) 2008-08-12 2010-02-18 Higuchi John W Non-Invasive Ocular Delivery of Rapamycin
US20100074863A1 (en) 2008-09-17 2010-03-25 Yat Sun Or Anti-infective pyrrolidine derivatives and analogs
AU2009293140A1 (en) 2008-09-17 2010-03-25 Endocyte, Inc. Folate receptor binding conjugates of antifolates
EP2174947A1 (en) 2008-09-25 2010-04-14 Universität des Saarlandes Bioactive pre-tubulysins and use thereof
WO2010045584A1 (en) 2008-10-17 2010-04-22 Endocyte, Inc. Folate targeting of nucleotides
WO2010045598A2 (en) 2008-10-17 2010-04-22 Purdue Research Foundation Psma binding ligand-linker conjugates and methods for using
IT1394860B1 (it) 2009-07-22 2012-07-20 Kemotech S R L Composti farmaceutici
US8394922B2 (en) 2009-08-03 2013-03-12 Medarex, Inc. Antiproliferative compounds, conjugates thereof, methods therefor, and uses thereof
ES2581207T3 (es) 2009-11-12 2016-09-02 Tube Pharmaceuticals Gmbh Inhibidores de la tubulina
EP2322537A1 (en) 2009-11-12 2011-05-18 R & D Biopharmaceuticals Gmbh Tubulin inhibitors
WO2011069116A1 (en) 2009-12-04 2011-06-09 Endocyte, Inc. Binding ligand linked drug delivery conjugates of tubulysins
US20120258905A1 (en) 2009-12-23 2012-10-11 Endocyte, Inc. Vitamin receptor drug delivery conjugates for treating inflammation
CA2790577A1 (en) * 2010-02-25 2011-09-01 Purdue Research Foundation Psma binding ligand-linker conjugates and methods for using
DK2528625T3 (da) 2010-04-15 2013-10-14 Spirogen Sarl Pyrrolobenzodiazepiner og konjugater deraf
EP2409983A1 (en) 2010-07-19 2012-01-25 Leibniz-Institut für Pflanzenbiochemie (IPB) Tubulysin analogues
AU2011285532B2 (en) * 2010-08-06 2015-09-10 Endocyte, Inc. Processes for preparing tubulysins
WO2012047525A2 (en) 2010-09-27 2012-04-12 Endocyte, Inc. Folate conjugates for treating inflammation of the eye
MX2013011201A (es) 2011-03-29 2013-12-16 Immunogen Inc Proceso para la elaboracion de conjugados de mejor homogeneidad.
RS58367B1 (sr) 2011-03-29 2019-03-29 Immunogen Inc Priprema konjugata antitela i majtanzinoida jednostepenim postupkom
CN103648532A (zh) 2011-06-21 2014-03-19 伊缪诺金公司 具有肽连接子的新型美登木素生物碱衍生物及其偶联物
CN104244718A (zh) 2011-12-05 2014-12-24 伊格尼卡生物治疗公司 抗体-药物缀合物以及相关化合物、组合物和方法
US10080805B2 (en) 2012-02-24 2018-09-25 Purdue Research Foundation Cholecystokinin B receptor targeting for imaging and therapy
EP2822386B1 (en) 2012-02-29 2021-05-05 Purdue Research Foundation Folate receptor alpha binding ligands
US20140080175A1 (en) 2012-03-29 2014-03-20 Endocyte, Inc. Processes for preparing tubulysin derivatives and conjugates thereof
CA2873112A1 (en) 2012-05-11 2013-11-14 Alexander Krantz Site-specific labeling and targeted delivery of proteins for the treatment of cancer
US9981046B2 (en) 2012-05-15 2018-05-29 Concortis Biosystems, Corp., a wholly owned Subsidiary of Sorrento Therapeutics, Inc. Drug-conjugates, conjugation methods, and uses thereof
PL2872157T3 (pl) 2012-07-12 2020-07-13 Hangzhou Dac Biotech Co., Ltd Koniugaty wiążących komórkę cząsteczek ze środkami cytotoksycznymi
EP2708243A1 (en) 2012-09-17 2014-03-19 OntoChem GmbH Receptor ligand linked cytotoxic molecules
US20140107316A1 (en) 2012-10-16 2014-04-17 Endocyte, Inc. Drug delivery conjugates containing unnatural amino acids and methods for using
EP2908818A4 (en) 2012-10-16 2016-07-13 Endocyte Inc CONJUGATES OF DRUG DELIVERY CONTAINING ARTIFICIAL AMINO ACIDS AND METHODS OF USE
BR112015011118B1 (pt) 2012-11-15 2022-12-13 Endocyte, Inc Conjugado; composição farmacêutica; e uso de um conjugado
US10131682B2 (en) 2012-11-24 2018-11-20 Hangzhou Dac Biotech Co., Ltd. Hydrophilic linkers and their uses for conjugation of drugs to a cell binding molecules
RU2015129800A (ru) 2012-12-21 2017-01-30 Биоэллаенс К. В. Гидрофильные саморазрушающиеся линкеры и их конъюгаты
RS56169B1 (sr) 2013-02-14 2017-11-30 Bristol Myers Squibb Co Jedinjenja tubulisina, metode pripreme i primena
US20140249315A1 (en) 2013-03-01 2014-09-04 Endocyte, Inc. Processes for preparing tubulysins
CN104784699B (zh) 2014-01-20 2019-05-03 博瑞生物医药（苏州）股份有限公司 叶酸受体结合配体-药物偶联物

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9550734B2 (en) 2004-07-23 2017-01-24 Endocyte, Inc. Bivalent linkers and conjugates thereof
US10647676B2 (en) 2004-07-23 2020-05-12 Endocyte, Inc. Bivalent linkers and conjugates thereof
US9555139B2 (en) 2007-03-14 2017-01-31 Endocyte, Inc. Binding ligand linked drug delivery conjugates of tubulysins
US10500204B2 (en) 2007-06-25 2019-12-10 Endocyte, Inc. Vitamin receptor drug delivery conjugates for treating inflammation
US10738086B2 (en) 2007-06-25 2020-08-11 Endocyte Inc. Conjugates containing hydrophilic spacer linkers
US9877965B2 (en) 2007-06-25 2018-01-30 Endocyte, Inc. Vitamin receptor drug delivery conjugates for treating inflammation
US10080805B2 (en) 2012-02-24 2018-09-25 Purdue Research Foundation Cholecystokinin B receptor targeting for imaging and therapy
US10765756B2 (en) 2012-02-24 2020-09-08 Purdue Research Foundation Cholecystokinin B receptor targeting for imaging and therapy
US11344623B2 (en) 2012-02-24 2022-05-31 Purdue Research Foundation Cholecystokinin B receptor targeting for imaging and therapy
US12485179B2 (en) 2012-02-24 2025-12-02 Purdue Research Foundation Cholecystokinin B receptor targeting for imaging and therapy
US9505747B2 (en) 2012-03-29 2016-11-29 Endocyte, Inc. Processes for preparing tubulysin derivatives and conjugates thereof
US9662402B2 (en) 2012-10-16 2017-05-30 Endocyte, Inc. Drug delivery conjugates containing unnatural amino acids and methods for using
WO2017031209A1 (en) * 2015-08-18 2017-02-23 Endocyte, Inc. Tubulysin analogs and methods
US11229708B2 (en) 2015-12-04 2022-01-25 Seagen Inc. Conjugates of quaternized tubulysin compounds
US11793880B2 (en) 2015-12-04 2023-10-24 Seagen Inc. Conjugates of quaternized tubulysin compounds

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