Formulation of a conjugate of a tubulysin analog to a cell-binding molecule

A formulation which is a liquid composition prior to lyophilization, or in a formulated lyophilized solid, or a reconstituted formulation from a lyophilized solid, comprising:

• a conjugate of a tubulysin compound to a cell-binding agent of Formula (I) or a pharmaceutically acceptable salt, hydrate, or hydrated salt thereof;
• or a polymorphic crystalline structure thereof;
• or an isotope, optical isomer, racemate, diastereomer or enantiomer thereof in an amount of 0.01%-99% by weight as a major ingredient;
• 0.0%-20.0% of one or more polyols;
• 0.0%-2.0% of one or more surfactants;
• 0.0%-5.0% of one or more preservatives;
• 0.0%-30% of one or more amino acids;
• 0.0%-5.0% of one or more antioxidants;
• 0.0%-0.3% of one or more metal chelating agents;
• 0.0%-30.0% of one or more buffer salts for adjusting pH of the formulation to pH 4.5 to 8.5;
• and 0.0%-30.0% of one or more of isotonic agents for adjusting osmotic pressure between about 250 to 350 mOsm after reconstituted for administration to a patient,
• wherein T is a cell-binding agent;
• L is a releasable linker;
• is a linkage bond that L connects to an atom inside the bracket independently;
• n is 1;
• and m is 1-10;
• wherein T is an antibody;
• a single chain antibody;
• an antibody fragment that binds to the target cell;
• a monoclonal antibody;
• a single chain monoclonal antibody;
• or a monoclonal antibody fragment that binds the target cell;
• a chimeric antibody;
• a chimeric antibody fragment that binds to the target cell;
• a domain antibody;
• a domain antibody fragment that binds to the target cell;
• wherein the linker L has a formula:
• —Ww-(Aa)r--Vv-;
• wherein:
• —W— is a Stretcher unit;
• w is 0 or 1;
• each --Aa-- is independently an amino acid unit;
• r is independently an integer ranging from 0 to 12;
• —V— is a Spacer unit;
• and v is 0, 1 or 2;
• the Stretcher unit W may independently contain a self-immolative spacer, a peptide unit, a hydrazone bond, a disulfide or thioether bond;
• the Stretcher unit (—W—), when present, links a targeted binding molecular unit (T) to the amino acid unit (--Aa--), or links V when an Aa is not present;
• W linked to T has a structure of:
• wherein R20 is selected from —C1-C9 alkylene-, —C1-C7 carbocyclo-, —O—(C1-C8 alkyl)-, -arylene-, —C1-C9 alkylene-arylene-, -arylene, —C1-C9 alkylene-, —C1-C9 alkylene-(C1-C8 carbocyclo)-, —(C3-C7 carbocyclo)-C1-C9 alkylene-, —C3-C8 heterocyclo-, —C1-C10 alkylene-(C3-C8 heterocyclo)-, —(C3-C8 heterocyclo)-C1-C9 alkylene-, —(CH2CH2O)k—, —(CH(CH3)CH2O)k—, and —(CH2CH2O)k—CH2—;
• k is an integer ranging from 1-20;
• the Spacer unit (—V—), when present, links an amino acid unit to an antimitotic agent when the amino acid unit is present;
• or, the Spacer unit links the Stretcher unit to an antimitotic agent when the amino acid unit is absent;
• or the Spacer unit links an antimitotic agent to the binding molecule (T) when both the amino acid unit and Stretcher unit are absent;
• the Spacer unit may contain one or more function groups that substantially increase water solubility, biological transport, preferential renal clearance, uptake, absorption, biodistribution, and/or bioavailability of the conjugate;
• the Spacer unit is self-immolative or non-self-immolative;
• a non-self-immolative Spacer unit is one in which part or all of the Spacer unit remains bound to the antimitotic agent after cleavage of the amino acid unit from the antimitotic agent-Linker-binding molecule conjugate or the antimitotic agent-Linker Compound;
• the self-immolative unit includes an aromatic compound that is electronically similar to a para-aminobenzyl-carbamoyl (PAB) group, 2-aminoimidazol-5-methanol compound, heterocyclic PAB compound, beta-glucuronide, and ortho or para-aminobenzylacetal;
• or one of following structures:
• wherein an (*) atom is a point of attachment of additional spacer or releasable linker unit(s), the antimitotic agent, and/or the cell binding agent (T);
• X, Y and Z3 are independently NH, O, or S;
• Z2 is H, NH, O or S;
• v is 0 or 1;
• Q is independently H, OH, C1-C6 alkyl, (OCH2CH2)nF, Cl, Br, I, OR17, SR17, NR17R18, N═NR17, N═R17, NR17R18, NO2, SOR17R18, SO2R17, SO3R17, OSO3R17, PR17R18, POR17R18, PO2R17R18, OPO(OR17)(OR18), or OCH2PO(OR17(OR18), wherein R17 and R18 are independently H, C1-C8 alkyl;
• C2-C8 alkenyl, alkynyl, or heteroalkyl;
• C3-C8 aryl, heterocyclic, carbocyclic, cycloalkyl, heterocycloalkyl, heteroaralkyl, or alkylcarbonyl;
• or a pharmaceutical cation salt thereof;
• the non-self-immolative spacer linker has a structure:
• or L- or D-natural or unnatural peptide containing 1-20 same or different amino acids;
• wherein “*” and “” atoms are a point of attachment of additional spacer or releasable linker, the antimitotic agent, and/or the cell-binding molecule;
• m is 1-10;
• n is 1-20;
• X2, X3, X4, X5, or X6, are independently selected from NH;
• NHNH;
• N(R12);
• N(R12)N(R12′);
• O;
• S;
• C1-C6 alkyl;
• C2-C6 heteroalkyl, alkylcycloalkyl, or heterocycloalkyl;
• C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl;
• CH2OR12, CH2SR12, CH2NHR12, or 1-8 amino acids;
• wherein R12 and R12′ are independently H;
• C1-C8 alkyl;
• C2-C8 hetero-alkyl, alkylcycloalkyl, or heterocycloalkyl;
• C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, alkylcarbonyl, or heteroaryl;
• or C1-C8 ester, ether, or amide;
• or a polyethyleneoxy unit of formula (OCH2CH2)p or (OCH2CH(CH3))p, wherein p is an integer from 0 to about 1000, or a combination thereof;
• a releasable component of the linker L that at least one bond in L can be broken under physiological conditions:
• a pH-labile, acid-labile, base-labile, oxidatively labile, metabolically labile, biochemically labile or enzyme-labile bond, which has one of following structures:
• —(CR15R16)m(Aa)r(CR17R18)n(OCH2CH2)t—, —(CR15R16)m(CR17R18)n(Aa)r(OCH2CH2)t—, -(Aa)r-(CR15R16)m(CR17R18)n(OCH2CH2)t—, —(CR15R16)m(CR17R18)n(OCH2CH2)(Aa)t-, —(CR15R16)m(CR17═CR18)(CR19R20)n(Aa)t(OCH2CH2)r—, —(CR15R16)m(NR11CO)(Aa)t(CR19R20)n—(OCH2CH2)r—, —(CR15R16)m(Aa)t(NR21CO)(CR19R20)n(OCH2CH2)r—, —(CR15R16)m(OCO)(Aa)t-(CR19R20)n(OCH2CH2)r—, —(CR15R16)m(OCNR17)(Aa)t(CR19R20)n(OCH2CH2)r—, —(CR15R16)m—(CO)(Aa)t-(CR19R20)n(OCH2CH2)r—, —(CR15R16)m(NR21CO)(Aa)t(CR19R20)n(OCH2CH2)r-, —(CR15R16)m—(OCO)(Aa)t(CR19R20)n—(OCH2CH2)r—, —(CR15R16)m(OCNR17)(Aa)t(CR19R20)n—(OCH2CH2)r—, —(CR15R16)m(CO)(Aa)t(CR19R20)n—(OCH2CH2)r—, —(CR15R16)m-phenyl-CO(Aa)t-(CR17R18)n—, —(CR15R16)m-furyl-CO(Aa)t(CR17R18)n—, —(CR15R6)m-oxazolyl-CO(Aa)t(CR17R18)n—, —(CR15R16)m-thiazolyl-CO(Aa)t(CCR17R18)n—, —(CR15R16)t-thienyl-CO(CR17R18)n—, —(CR15R16)t-imidazolyl-CO—(CR17R18)n—, —(CR15R16)t-morpholino-CO (Aa)t-(CR17R18)n—, —(CR15R16)t-piperazino-CO(Aa)t(CR17R18)n—, —(CR15R16)t—N-methylpiperazin-CO(Aa)t(CR17R18)n—, —(CR15R16)m-(Aa)phenyl-, —(CR15R16)m-(Aa)tfuryl-, —(CR15R16)m-oxazolyl(Aa)t-, —(CR15R16)m-thiazolyl(Aa)t-, —(CR15R16)m-thienyl-(Aa)t-, —(CR15R16)m-imidazolyl(Aa)t-, —(CR15R16)m-morpholino-(Aa)t-, —(CR15R16)m-piperazino-(Aa)t-, —(CR15R16)m-N-methylpiperazino-(Aa)t-, —K(CR15R16)m(Aa)r(CR17R18)n(OCH2CH2)t—, —K(CR15R16)m(CR17R18)n(Aa)r(OCH2CH2)t—, —K(Aa)r-(CR15R16)m(CR17R18)n(OCH2CH2)t—, —K(CR15R16)m(CR17R18)n(OCH2CH2)r(Aa)t-, —K(CR15R16)m—(CR17═CR18)(CR19R20)n(Aa)t(OCH2CH2)r, —K(CR15R16)m(NR11CO)(Aa)t-(CR19R20)n(OCH2CH2)r—, —K(CR5R6)m(Aa)t(NR21CO)(CR19R20)n(OCH2CH2)r—, —K(CR15R16)m—(OCO)(Aa)t(CR19R20)n—(OCH2CH2)r—, —K(CR15R16)m(OCNR17)(Aa)t(CR19R20)n—(OCH2CH2)r—, —K(CR15R16)m(CO)(Aa)t- (CR19R20)n(OCH2CH2)r—, —K(CR15R16)m(NR21CO)(Aa)t-(CR19R20)n—(OCH2CH2)r—, —K(CR15R16)m—(OCO)(Aa)t(CR19R20)n(OCH2CH2)r—, —K(CR15R16)m—(OCNR17)(Aa)t-(CR19R20)n(OCH2CH2)r—, —K—(CR15R16)m(CO)(Aa)t(CR19R20)n(OCH2CH2)r—, —K(CR15R16)m-phenyl-CO(Aa)t(CR17R18)n—, —K—(CR15R16)m-furyl-CO(Aa)t(CR17R18)n—, —K(CR15R16)m-oxazolyl-CO(Aa)t(CR17R18)n—, —K(CR15R16)m-thiazolyl-CO(Aa)t-(CR17R18)n—, —K(CR15R16)t-thienyl-CO(CR17R18)n—, —K(CR15R16)timidazolyl-CO—(CR17R18)n—, —K(CR5R6)tmorpholino-CO(Aa)t-(CR17R18)n—, —K(CR15R16)t-piperazino-CO(Aa)t-(CR17R18)n—, —K(CR15R16)t—N-methylpiperazin-CO(Aa)t(CR17R18)n—, —K(CR15R16)m-(Aa)phenyl, —K—(CR15R16)m-(Aa)tfuryl-, —K(CR15R16)m-oxazolyl-(Aa)t-, —K(CR15R16)m-thiazolyl(Aa)t-, —K(CR15R16)m-thienyl-(Aa)t-, —K(CR15R16)m-imidazolyl(Aa)t-, —K(CR15R16)m-morpholino(Aa)t-, —K(CR15R16)mpiperazino(Aa)tG, —K(CR5R6)m—N-methyl-piperazino(Aa)t-;
• wherein m, Aa, m, n, R13, R14, and R15 are described above;
• t and r are 0-100 independently;
• R16, R17, R18, R19, and R20 are independently chosen from H;
• halide;
• C1-C8alkyl or heteroalkyl, C2-C8 aryl, alkenyl, alkynyl, ether, ester, amine or amide, C3-C8 aryl, which is optionally substituted by one or more halide, CN, NR12R12′, CF3, OR12, aryl, heterocycle, S(O)R12, SO2R12, —CO2H, —SO3H, —OR12, —CO2R12, —CONR12, —PO2R12R13, —PO3H or P(O)R12R12′R13;
• K is NR12, —SS—, —C(═O)—, —C(═O)NH—, —C(═O)O—, —C═NH—O—, —C═N—NH—, —C(═O)NH—NH—, O, S, Se, B, Het (heterocyclic or heteroaromatic ring having C3-C12);
• or a peptide containing same or different 1-20 amino acids;
• inside the bracket of the Formula (I) is a tubulysin compound wherein R1, R2, R3, and R4 are independently absent, a linear or branched C1-C8alkyl, or alkyl alcohol;
• C2-C8 heteroalkyl, alkylcycloalkyl, heterocycloalkyl, alkyl ether, alkyl carboxylate, alkyl amine, alkyl ester, or alkyl amide;
• C3-C8 aryl, Ar-alkyl, heterocyclic, carbocyclic, cycloalkyl, heteroalkylcycloalkyl, or alkylcarbonyl;
• or two R's:
• R1 and R2, R3 and R4, R5 and R6, or R12 and R13 together form a 3-7 membered carbocyclic, cycloalkyl, heterocyclic, heterocycloalkyl, aromatic or heteroaromatic ring system;
• Y is N or C;
• wherein R5, R6, R8 and R10 are independently selected from H and a linear or branched C1-C4 alkyl or C2-C4 of heteroalkyl;
• wherein R7is selected from H, R14, or —R14C(═O)X1R15;
• —R14X1R15, X1 is O, S, S—S, NH, or NR14;
• wherein R9 is H, —O—, —OR14, —OC(═O)R14—, —OC(═O)NHR14—, —OC(═O)NR14R15—, —OC(═O)R14SSR15—, OP(═O)(OR14)—, or OR14OP(═O)(OR15);
• wherein R11 is H, R14, —R14C(═O)R16, —R14C(═O)X2R16, —R14X2R16, or —R14C(═O)X2, wherein X2 is —O—, —S—, —NH—, —NHS(O2), —NHS(O), —N(R14)—, —O—R14—, —S—R14—, —S(═O)—R14—, or —NHR14—;
• wherein R12 is H, R14, —O—, —S—, —N—, ═N—, ═NNH—, —OH, —SH, —NH2, ═NH, ═NNH2, —NH(R14), —OR14, —C(O)O—, —C(O)OR16—, —COR16, —COOR14—, C(O)NH—, C(O)NH2, C(O)NHR14, —SR14, —S(═O)R14, —P(═O)(OR16)2, —OP(═O)(OR16)2, —CH2OP(═O)(OR16)2, or —SO2R16;
• wherein R13 is a linear or branched C1-C10 alkyl, alkyl acid, alkyl amide, or alkyl amine;
• or C2-C10 heteroalkyl;
• or C3-C10 Ar;
• Ar is an aromatic or hetero aromatic group, composed of one or several rings, comprising four to ten carbon atoms, wherein one or several H atoms in the aromatic group can optionally be replaced independently by R17, F, Cl, Br, I, OR16, SR16, NR16R17, N═NR16, N═R16, NR16R17, NO2, SOR16R17, SO2R16, SO3R16, OSO3R16, PR16R17, POR16R17, PO2R16R17, OP(O)(OR17)2, OCH2OP(O)(OR17)2, OC(O)OP(O)(OR17)2, PO(OR16)(OR17), OP(O)(OR17)OP(O)(OR17)2, OC(O)R17 or OC(O)NHR17;
• wherein R14 and R15 are independently H;
• a linear or branched C1-C8 alkyl;
• C2-C8 alkenyl, alkynyl, heteroalkyl, heterocyclic, or carbocyclic;
• or C3-C8 aryl, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroaralkyl, heteroalkylcycloalkyl, or alkylcarbonyl;
• wherein when R14 is bivalent, R14 is further connected to an additional functional group of one to four amino acid units, or (CH2CH2O)r, r is an integer ranging from 0 to 12, or C4-C12 glycoside, or C1-C8 carboxylic acid;
• wherein R16 is H, OH, R14 or one to four amino acid units;
• wherein R17 is H, a linear or branched C1-C8 alkyl;
• C2-C8 alkenyl, alkynyl, heteroalkyl, or heterocyclic;
• C3-C8 aryl, carbocyclic, cycloalkyl, alkylcycloalkyl, heterocycloalkyl, heteroalkylcycloalkyl, heteroaralkyl, or alkylcarbonyl;
• or C4-C12 glycoside, or a pharmaceutical salt thereof;
• wherein the formulation further contains a buffer agent.