# US 11,998,611 B2 · Cryptophycin antibody conjugates for the treatment of cancer

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- Number: US 11,998,611 B2
- Document ID: US11998611B2
- Status: Granted
- Kind: B2
- Application number: 17/210,072
- Filed: 2021-03-23
- Published: 2021-07-29
- Granted: 2024-06-04
- Earliest claimed: 2017-05-10
- Assignees: Sanofi
- Applicants: Sanofi
- Inventors: Hervé Bouchard; Marie-Priscille Brun; Philippe Hubert
- Web page: https://patentgrep.com/patent/US11998611B2

## CPC

- A61K 47/65: Peptidic linkers, binders or spacers, e.g. peptidic enzyme-labile linkers
- A61K 38/15: Depsipeptides; Derivatives thereof
- A61K 47/10: Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
- A61K 47/22: Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
- A61K 47/64: Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent (peptidic linkers A61K47/65)
- A61K 47/6829: {Bacterial toxins, e.g. diphteria toxins or Pseudomonas exotoxin A}
- A61K 47/6889: {Conjugates wherein the antibody being the modifying agent and wherein the linker, binder or spacer confers particular properties to the conjugates, e.g. peptidic enzyme-labile linkers or acid-labile linkers, providing for an acid-labile immuno conjugate wherein the drug may be released from its antibody conjugated part in an acidic, e.g. tumoural or environment}
- A61P 35/00: Antineoplastic agents
- C07D 405/06: linked by a carbon chain containing only aliphatic carbon atoms
- C07D 405/14: containing three or more hetero rings
- C07K 11/00: Depsipeptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present disclosure relates, inter alia, to cryptophycin conjugates of formula (V): The disclosure also relates to precursors of these conjugates, compositions containing these conjugates, and to their therapeutic use, especially as anticancer agents. The disclosure also relates to the process for preparing these conjugates.

## Claims

### Claim 1

A compound of formula (V):
wherein:
R1 is (C1-C6)alkyl;
R2 is hydrogen or (C1-C6)alkyl;
R3 is hydrogen or (C1-C6)alkyl;
R4 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, (C1-C6)alkylene-NHR12, (C1-C6)alkylene-OH, or (C1-C6)alkylene-SH;
R5 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, (C1-C6)alkylene-NHR12, (C1-C6)alkylene-OH, or (C1-C6)alkylene-SH;
X is —NR6— or —O—;
R6 is hydrogen or (C1-C6)alkyl;
R7 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, or (C1-C6)alkylene-N[(C1-C6)alkyl]2;
R8 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, or (C1-C6)alkylene-N[(C1-C6)alkyl]2;
each R9 is independently hydrogen, halogen, NH2, NH(C1-C6)alkyl, N[(C1-C6)alkyl]2, NH(C3-C6)cycloalkyl, OH, O(C1-C4)alkyl, or (C3-C6)heterocycloalkyl;
each R10 is independently hydrogen or (C1-C4)alkyl;
each R12 is independently hydrogen or (C1-C6)alkyl;
Y is —NR11—(C1-C6)alkylene-, —O—(C1-C6)alkylene-, or —S—(C1-C6)alkylene-, wherein the (C1-C6)alkylene of Y is bonded to the phenyl ring and the —NR11—, —O—, or —S— of Y is either (a) bonded directly to the —C(O)— of L1 in formula (II) or (b) bonded directly to the —C(O)— of an (AA)w of L1 in formula (II);
R11 is hydrogen or (C1-C6)alkyl; and
L is formula (II):
wherein:
L1 is formula (III):
wherein:
(i) x is 0;
y is 0; and
z is 0; or
(ii) x is 1;
y is 0; and
z is 0; or
(iii) x is 1;
y is 1; and
z is 1;
A5 is hydrogen or (C1-C6)alkyl;
ALK is (C1-C12)alkylene;
A6 is hydrogen or (C1-C6)alkyl;
A2 is hydrogen or (C1-C6)alkyl;
A3 is hydrogen or (C1-C6)alkyl;
J1 is CA1 or N;
J2 is CA1 or N;
J3 is CA1 or N;
J4 is CA1 or N;
each A1 is independently hydrogen or (C1-C6)alkyl; and
A4 is hydrogen or (C1-C6)alkyl;
wherein L1 is bonded directly to —O—, or —S— of Y;
(AA)w is a sequence of w amino acids connected together via peptide bonds, wherein each amino acid, (AA) W, is independently a natural or non-natural amino acid, (AAns);
w is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
each (AA n O is independently selected from the group consisting of alanine (Ala), γ-aminobutyric acid (GABA), arginine (Arg), asparagine (Asn), aspartic acid (Asp), citrulline (Cit), cysteine (Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), ε-(acetyl)-lysine ((ε-Ac)Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val); and
L2 is:
(a) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(b) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(c) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(d) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(e) —NA7-(C1-C6)alkylene-;
(f) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(g) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(h) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(i) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(j) —C(O)—NA7-(C1-C6)alkylene-;
(k) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(l) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(m) —C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(n) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(o) —(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(p) —C(O)—(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(q) —NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(r) —C(O)—NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(s) —C(O)—NA7-(OCH2CH2)i—(C1-C6)alkylene-;
(t) —NA7-(CH2CH2O)i—(C1-C6)alkylene-;
(u) —(C1-C6)alkylene-NA7-;
(v) —C(O)—(C1-C6)alkylene-NA7-;
(w) —NA8-(C1-C6)alkylene-NA7-;
(x) —C(O)—NA8-(C1-C6)alkylene-NA7-;
(y) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(z) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(aa) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(bb) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(cc) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(dd) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ee) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ff) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(gg) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(hh) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ii) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(jj) —C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(kk) —NA7-arylene-;
(ll) —C(O)—NA7-arylene-;
(mm) —NA7-heteroarylene-; or
(nn) —C(O)—NA7-heteroarylene-;
A7 is a straight, branched, saturated, or unsaturated C1-C160 hydrocarbon;
wherein one or more —CH2— units of the C1-C160 hydrocarbon are optionally and independently replaced by one or more atoms or groups independently selected from the group consisting of —CH(OH)—, —CH(SO3H)—, —CH(SO3−M+)—, —C(O)—, —C(O)NH—, —C(O)N(alkyl)-, —C(O)O—, —NHC(O)—, —N(alkyl)C(O)—, —NHS(O)2—, —N(alkyl)S(O)2—, —P(O)(OH)—, —P(O)(OH)O—, —O—, —OC(O)—, —OC(O)O—, —OP(O)(OH)—, —OP(O)(OH)O—, —S(O)—, —S(O)2—, —S(O)2NH—, —S(O)2N(alkyl)-, and heterocycloalkyl;
wherein each heterocycloalkyl is optionally and independently substituted by one or more substituents independently selected from the group consisting of halogen, alkyl, NH2, NH(alkyl), N(alkyl)2, OH, and O(alkyl); and
wherein the C1-C160 hydrocarbon is optionally substituted by one or more substituents independently selected from the group consisting of halogen, alkyl, NH2, NH(alkyl), N(alkyl)2, OH, and O(alkyl);
A8 is hydrogen or (C1-C6)alkyl;
M+ is an alkali metal; and
i is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, or 50;
wherein the right end of L2 is bonded to —C(O)— and the left end of L2 is bonded to RCG1 of G;
G is the product of a reaction between RCG1, a reactive chemical group present at the end of L of formula (II), and RCG2, an orthogonal reactive chemical group present on an antibody, Ab;
(i) RCG1 is:
Cl,C≡CH,NH2,N3,OH,O(alkyl)-NHOH,SH,
wherein:
R21 is hydrogen or (C1-C6) alkyl; and
the —CO—, —NH—, —NR21—, —N═, —O—, or S is bonded to L2; or
(ii) RCG1 is:
C(O)ZaRa,
wherein:
Za is a single bond, —NH—, or —O—; and
Ra is hydrogen, (C1-C6)alkyl, alkenyl, (C3-C7)cycloalkyl, (C3-C7)heterocycloalkyl, aryl, or heteroaryl, wherein the (C3-C7)heterocycloalkyl, aryl, or heteroaryl is optionally substituted by 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of halogen, CN, NO2, alkyl, OH, O(alkyl), and =0;
wherein —C(O)— is bonded to L2;
RCG2 is:
C≡CH,C(O)H,C(O)C1-C6alkyl,C(O)NH2,C(O)OH,NH2,N3,SH,
wherein:
R21 is hydrogen or (C1-C6) alkyl; and
Ab is the antibody.

### Claim 2 (depends on 1)

The compound according to claim 1, wherein the compound has the following structure:

### Claim 3 (depends on 1)

The compound according to claim 1, wherein (AA) w is Ala-Lys, Ala-Phe, Gly-Gly, Ile-Cit, Leu-Cit, Phe-Ala, Phe-Cit, Phe-Gly, Phe-Lys, Trp-Cit, Val-Ala, Val-Lys, Val-(ε-Ac)Lys, Val-Cit, Gly-Ala-Phe, Gly-Gly-Gly, Gly-Phe-Gly, Gly-Phe-Lys, Gly-Val-Cit, Phe-Phe-Lys, D-Phe-Phe-Lys, Ala-Leu-Ala-Leu, Gly-Phe-Leu-Cit, or Gly-Phe-Leu-Gly.

### Claim 4 (depends on 1)

The compound according to claim 1, wherein L2 is:
(a) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(b) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(c) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(d) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(e) —NA7-(C1-C6)alkylene-;
(f) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(g) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(h) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(i) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(k) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(l) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(m) —C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(n) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(o) —(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(p) —C(O)—(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(q) —NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(r) —C(O)—NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(s) —C(O)—NA7-(OCH2CH2)i—(C1-C6)alkylene-;
(t) —NA7-(CH2CH2O)i—(C1-C6)alkylene-;
(u) —(C1-C6)alkylene-NA7-;
(v) —C(O)—(C1-C6)alkylene-NA7-;
(w) —NA8-(C1-C6)alkylene-NA7-;
(x) —C(O)—NA8-(C1-C6)alkylene-NA7-;
(y) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(z) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(aa) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(bb) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(cc) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(dd) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ee) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ff) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(gg) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(hh) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ii) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(jj) —C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(kk) —NA7-arylene-;
(ll) —C(O)—NA7-arylene-;
(mm) —NA7-heteroarylene-; or
(nn) —C(O)—NA7-heteroarylene-.

### Claim 5 (depends on 1)

The compound according to claim 1, wherein L2 is selected from the group consisting of:

### Claim 6 (depends on 1)

The compound according to claim 1, wherein A7 is C(O)—[CH2CH2O]4—CH3, C(O)—[CH2CH2O]7—CH3, C(O)—[CH2CH2O]24—CH3, C(O)CH2CH2C(O)NHCH2CH2S(O)2OH, C(O)CH2CH2C(O)NHCH2CH2S(O)2OM+, C(O)CH2CH2C(O)NH—[CH2CH2O]4—CH2CH2C(O)NHCH2CH2S(O)2OH, or C(O)CH2CH2C(O)NH—[CH2CH2O]4—CH2CH2C(O)NHCH2CH2S(O)2OM+.

### Claim 7 (depends on 1)

The compound according to claim 1, wherein:
L2 is —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
A7 is C(O)CH2CH2C(O)NHCH2CH2S(O)2OH, C(O)CH2CH2C(O)NHCH2CH2S(O)2OM+, C(O)CH2CH2C(O)NH—[CH2CH2O]a—CH2CH2C(O)NHCH2CH2S(O)2OH, C(O)CH2CH2C(O)NH—[CH2CH2O]a—CH2CH2C(O)NHCH2CH2S(O)2OM+, or C(O)—[CH2CH2O]a—CH3; and
a is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, or 50.

### Claim 8 (depends on 1)

The compound according to claim 1, wherein L is selected from the group consisting of:

### Claim 9 (depends on 1)

The compound according to claim 1, wherein RCG1 is C(O)ZaRa, wherein —C(O)— is bonded to L2.

### Claim 10 (depends on 1)

The compound according to claim 1, wherein RCG2 is selected from the group consisting of:
(i) a C(O)H group of an engineered formylglycine residue of the antibody;
(ii) a C(O)NH2 group borne by the side chain of a glutamine residue that is present at the surface of the antibody;
(iii) an α-NH2 group of an N-terminal amino acid of a heavy chain or a light chain of the antibody;
(iv) an ε-NH2 group borne by the side chain of a lysine residue that is present at the surface of the antibody;
(v) an SH of a cysteine residue generated by reducing an intra-chain disulfide bond of the antibody;
(vi) an SH of an engineered cysteine residue of the antibody; and
(vii) a saccharide group of the hinge region of the antibody.

### Claim 11 (depends on 1)

The compound according to claim 1, wherein:
(i) RCG1 is Cl,
and
RCG2 is SH; or
(ii) RCG1 is C≡CH,
and
RCG2 is N3; or
(iii) RCG1 is NH 2 or OH; and
RCG2 is C(O)NH2 or C(O)OH; or
(iv) RCG1 is N3; and
RCG2 is C≡CH,
or
(v) RCG1 is O(alkyl)-NHOH or
and
RCG2 is C(O)H or C(O)C1-C6 alkyl; or
(vi) RCG1 is SH; and
RCG2 is
or
(vii) RCG1 is
and
RCG2 is NH2.

### Claim 12 (depends on 1)

The compound according to claim 1, wherein G is selected from the group consisting of:

### Claim 13 (depends on 1)

The compound according to claim 1, wherein the compound is selected from the group consisting of:

### Claim 14 (depends on 1)

A medicament comprising a compound according to claim 1.

### Claim 15 (depends on 1)

A pharmaceutical composition comprising a compound according to claim 1 and a pharmaceutically acceptable excipient.

### Claim 16 (depends on 1)

A method for treating cancer in a patient in need thereof, wherein the method comprises administering to the patient a therapeutically effective amount of a compound according to claim 1.

### Claim 17 (depends on 1)

A process for preparing a compound of formula (V) according to claim 1:
wherein:
R1 is (C1-C6)alkyl;
R2 is hydrogen or (C1-C6)alkyl;
R3 is hydrogen or (C1-C6)alkyl;
R4 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, (C1-C6)alkylene-NHR12, (C1-C6)alkylene-OH, or (C1-C6)alkylene-SH;
R5 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, (C1-C6)alkylene-NHR12, (C1-C6)alkylene-OH, or (C1-C6)alkylene-SH;
X is —NR6— or —O—;
R6 is hydrogen or (C1-C6)alkyl;
R7 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, or (C1-C6)alkylene-N[(C1-C6)alkyl]2;
R8 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, or (C1-C6)alkylene-N[(C1-C6)alkyl]2;
each R9 is independently hydrogen, halogen, NH2, NH(C1-C6)alkyl, N[(C1-C6)alkyl]2, NH(C3-C6)cycloalkyl, OH, O(C1-C4)alkyl, or (C3-C6)heterocycloalkyl;
each R10 is independently hydrogen or (C1-C4)alkyl;
each R12 is independently hydrogen or (C1-C6)alkyl;
Y is —NR11—(C1-C6)alkylene-, —O—(C1-C6)alkylene-, or —S—(C1-C6)alkylene-, wherein the (C1-C6)alkylene of Y is bonded to the phenyl ring and the —NR11—, —O—, or —S— of Y is either (a) bonded directly to the —C(O)— of L1 in formula (II) or (b) bonded directly to the —C(O)— of an (AA)w of L1 in formula (II);
R11 is hydrogen or (C1-C6)alkyl; and
L is formula (II):
wherein:
L1 is formula (III):
wherein:
(i) x is 0;
y is 0; and
z is 0; or
(ii) x is 1;
y is 0; and
z is 0; or
(iii) x is 1;
y is 1; and
z is 1;
A5 is hydrogen or (C1-C6)alkyl;
ALK is (C1-C12)alkylene;
A6 is hydrogen or (C1-C6)alkyl;
A2 is hydrogen or (C1-C6)alkyl;
A3 is hydrogen or (C1-C6)alkyl;
J1 is CA1 or N;
J2 is CA1 or N;
J3 is CA1 or N;
J4 is CA1 or N;
each A1 is independently hydrogen or (C1-C6)alkyl; and
A4 is hydrogen or (C1-C6)alkyl;
wherein L1 is bonded directly to —NR11—, —O—, or —S— of Y;
(AA)w is a sequence of w amino acids connected together via peptide bonds, wherein each amino acid, (AA)w, is independently a natural or non-natural amino acid, (AAns);
w is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
each (AA n O is independently selected from the group consisting of alanine (Ala), γ-aminobutyric acid (GABA), arginine (Arg), asparagine (Asn), aspartic acid (Asp), citrulline (Cit), cysteine (Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), ε-(acetyl)-lysine ((ε-Ac)Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val); and
L2 is:
(a) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(b) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(c) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(d) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(e) —NA7-(C1-C6)alkylene-;
(f) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(g) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(h) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(i) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(j) —C(O)—NA7-(C1-C6)alkylene-;
(k) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(l) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(m) —C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(n) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(o) —(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(p) —C(O)—(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(q) —NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(r) —C(O)—NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(s) —C(O)—NA7-(OCH2CH2)i—(C1-C6)alkylene-;
(t) —NA7-(CH2CH2O)i—(C1-C6)alkylene-;
(u) —(C1-C6)alkylene-NA7-;
(v) —C(O)—(C1-C6)alkylene-NA7-;
(w) —NA8-(C1-C6)alkylene-NA7-;
(x) —C(O)—NA8-(C1-C6)alkylene-NA7-;
(y) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(z) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(aa) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(bb) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(cc) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(dd) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ee) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ff) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(gg) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(hh) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ii) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(jj) C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(kk) —NA7-arylene-;
(ll) —C(O)—NA7-arylene-;
(mm)-NA7-heteroarylene-; or
(nn) —C(O)—NAS-heteroarylene-;
A7 is a straight, branched, saturated, or unsaturated C1-C160 hydrocarbon;
wherein one or more —CH2— units of the C1-C160 hydrocarbon are optionally and independently replaced by one or more atoms or groups independently selected from the group consisting of —CH(OH)—, —CH(SO3H)—, —CH(SO3−M+)—, —C(O)—, —C(O)NH—, —C(O)N(alkyl)-, —C(O)O—, —NHC(O)—, —N(alkyl)C(O)—, —NHS(O)2—, —N(alkyl)S(O)2—, —P(O)(OH)—, —P(O)(OH)O—, —O—, —OC(O)—, —OC(O)O—, —OP(O)(OH)—, —OP(O)(OH)O—, —S(O)—, —S(O)2—, —S(O)2NH—, —S(O)2N(alkyl)-, and heterocycloalkyl;
wherein each heterocycloalkyl is optionally and independently substituted by one or more substituents independently selected from the group consisting of halogen, alkyl, NH2, NH(alkyl), N(alkyl)2, OH, and O(alkyl); and
wherein the C1-C160 hydrocarbon is optionally substituted by one or more substituents independently selected from the group consisting of halogen, alkyl, NH2, NH(alkyl), N(alkyl)2, OH, and O(alkyl);
A8 is hydrogen or (C1-C6)alkyl;
M+ is an alkali metal; and
i is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, or 50;
wherein the right end of L2 is bonded to —C(O)— and the left end of L2 is bonded to RCG1 of G;
G is the product of a reaction between RCG1, a reactive chemical group present at the end of L of formula (II), and RCG2, an orthogonal reactive chemical group present on an antibody, Ab;
(i) RCG1 is:
Cl,C≡CH,NH2,N3,OH,O(alkyl)-NHOH,SH,
wherein:
R21 is hydrogen or (C1-C6) alkyl; and
the —CO—, —NH—, —NR21—, —N═, —O—, or S is bonded to L2; or
(ii) RCG1 is:
C(O)ZaRa,
wherein:
Za is a single bond, —NH—, or —O—; and
Ra is hydrogen, (C1-C6)alkyl, alkenyl, (C3-C7)cycloalkyl, (C3-C7)heterocycloalkyl, aryl, or heteroaryl, wherein the (C3-C7)heterocycloalkyl, aryl, or heteroaryl is optionally substituted by 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of halogen, CN, NO2, alkyl, OH, O(alkyl), and ═O;
wherein —C(O)— is bonded to L2;
RCG2 is:
C≡CH,C(O)H,C(O)C1-C6 alkyl,C(O)NH2,C(O)OH,NH2,N3,SH,
wherein:
R21 is hydrogen or (C1-C6) alkyl; and
Ab is the antibody;
wherein the process comprises the following steps:
(i) contacting and reacting a solution of a compound of formula (IV):
wherein:
R1 is (C1-C6)alkyl;
R2 is hydrogen or (C1-C6)alkyl;
R3 is hydrogen or (C1-C6)alkyl;
R4 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, (C1-C6)alkylene-NHR12, (C1-C6)alkylene-OH, or (C1-C6)alkylene-SH;
R5 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, (C1-C6)alkylene-NHR12, (C1-C6)alkylene-OH, or (C1-C6)alkylene-SH;
X is —NR6— or —O—;
R6 is hydrogen or (C1-C6)alkyl;
R7 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, or (C1-C6)alkylene-N[(C1-C6)alkyl]2;
R8 is hydrogen, (C1-C6)alkyl, (C1-C6)alkylene-C(O)OH, or (C1-C6)alkylene-N[(C1-C6)alkyl]2;
each R9 is independently hydrogen, halogen, NH2, NH(C1-C6)alkyl, N[(C1-C6)alkyl]2, NH(C3-C6)cycloalkyl, OH, O(C1-C4)alkyl, or (C3-C6)heterocycloalkyl;
each R10 is independently hydrogen or (C1-C4)alkyl;
each R12 is independently hydrogen or (C1-C6)alkyl;
Y is —NR11—(C1-C6)alkylene-, —O—(C1-C6)alkylene-, or —S—(C1-C6)alkylene-, wherein the (C1-C6)alkylene of Y is bonded to the phenyl ring and the —NR11—, —O—, or —S— of Y is either (a) bonded directly to the —C(O)— of L1 in formula (II) or (b) bonded directly to the —C(O)— of an (AA)w of L1 in formula (II);
R11 is hydrogen or (C1-C6)alkyl; and
L is formula (II):
wherein:
L1 is formula (III):
wherein:
(i) x is 0;
y is 0; and
z is 0; or
(ii) x is 1;
y is 0; and
z is 0; or
(iii) x is 1;
y is 1; and
z is 1;
A5 is hydrogen or (C1-C6)alkyl;
ALK is (C1-C12)alkylene;
A6 is hydrogen or (C1-C6)alkyl;
A2 is hydrogen or (C1-C6)alkyl;
A3 is hydrogen or (C1-C6)alkyl;
J1 is CA1 or N;
J2 is CA1 or N;
J3 is CA1 or N;
J4 is CA1 or N;
each A1 is independently hydrogen or (C1-C6)alkyl; and
A4 is hydrogen or (C1-C6)alkyl;
wherein L1 is bonded directly to —NR11—, —O—, or —S— of Y;
(AA)w is a sequence of w amino acids connected together via peptide bonds, wherein each amino acid, (AA)w, is independently a natural or non-natural amino acid, (AAns);
w is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12;
each (AA n O is independently selected from the group consisting of alanine (Ala), γ-aminobutyric acid (GABA), arginine (Arg), asparagine (Asn), aspartic acid (Asp), citrulline (Cit), cysteine (Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), ε-(acetyl)-lysine ((ε-Ac)Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val); and
L2 is:
(a) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(b) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(c) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(d) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-;
(e) —NA7-(C1-C6)alkylene-;
(f) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(g) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(h) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(i) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-;
(j) —C(O)—NA7-(C1-C6)alkylene-;
(k) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(l) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(m) —C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(n) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-;
(o) —(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(p) —C(O)—(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(q) —NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(r) —C(O)—NA8-(C1-C6)alkylene-(OCH2CH2)i—NA7-(C1-C6)alkylene-;
(s) —C(O)—NA7-(OCH2CH2)i—(C1-C6)alkylene-;
(t) —NA7-(CH2CH2O)i—(C1-C6)alkylene-;
(u) —(C1-C6)alkylene-NA7-;
(v) —C(O)—(C1-C6)alkylene-NA7-;
(w) —NA8-(C1-C6)alkylene-NA7-;
(x) —C(O)—NA8-(C1-C6)alkylene-NA7-;
(y) —(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(z) —C(O)—(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(aa) —NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH2CH2)i—;
(bb) —C(O)—NA8-(C1-C6)alkylene-NA7-C(O)—(C1-C6)alkylene-(OCH 2CH2)i—;
(cc) —(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(dd) —C(O)—(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ee) —NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ff) —C(O)—NA8-(C1-C6)alkylene-NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(gg) —(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(hh) —C(O)—(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(ii) —NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
C(O)—NA8-(C1-C6)alkylene-C(O)—NA7-(C1-C6)alkylene-(OCH2CH2)i—;
(kk) —NA7-arylene-;
(ll) —C(O)—NA7-arylene-;
(mm)-NA7-heteroarylene-; or
(nn) —C(O)—NA7-heteroarylene-;
A7 is a straight, branched, saturated, or unsaturated C1-C160 hydrocarbon;
wherein one or more —CH2— units of the C1-C160 hydrocarbon are optionally and independently replaced by one or more atoms or groups independently selected from the group consisting of —CH(OH)—, —CH(SO3H)—, —CH(SO3−M+)—, —C(O)—, —C(O)NH—, —C(O)N(alkyl)-, —C(O)O—, —NHC(O)—, —N(alkyl)C(O)—, —NHS(O)2—, —N(alkyl)S(O)2—, —P(O)(OH)—, —P(O)(OH)O—, —O—, —OC(O)—, —OC(O)O—, —OP(O)(OH)—, —OP(O)(OH)O—, —S(O)—, —S(O)2—, —S(O)2NH—, —S(O)2N(alkyl)-, and heterocycloalkyl;
wherein each heterocycloalkyl is optionally and independently substituted by one or more substituents independently selected from the group consisting of halogen, alkyl, NH2, NH(alkyl), N(alkyl)2, OH, and O(alkyl); and
wherein the C1-C160 hydrocarbon is optionally substituted by one or more substituents independently selected from the group consisting of halogen, alkyl, NH2, NH(alkyl), N(alkyl)2, OH, and O(alkyl);
A8 is hydrogen or (C1-C6)alkyl;
M+ is an alkali metal; and
i is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44,
(i) RCG1 is:
Cl,C≡CH,NH2,N3,OH,O(alkyl)-NHOH,SH,
wherein:
R21 is hydrogen or (C1-C6) alkyl; and
the —CO—, —NH—, —NR21—, —N═, —O—, or S is bonded to L2; or
(ii) RCG1 is:
C(O)ZaRa,
wherein:
Za is a single bond, —NH—, or —O—; and
Ra is hydrogen, (C1-C6)alkyl, alkenyl, (C3-C7)cycloalkyl, (C3-C7)heterocycloalkyl, aryl, or heteroaryl, wherein the (C3-C7)heterocycloalkyl, aryl, or heteroaryl is optionally substituted by 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of halogen, CN, NO2, alkyl, OH, O(alkyl), and =0;
wherein —C(O)— is bonded to L2;
with an optionally buffered aqueous solution of an antibody, Ab, containing an orthogonal reactive chemical group, RCG2, present on the antibody, wherein for each RCG1, RCG2 is independently selected from the group consisting of:
C≡CH,C(O)H,C(O)C1-C6 alkyl,C(O)NH2,C(O)OH,NH2,N3,SH,
wherein:
R21 is hydrogen or (C1-C6) alkyl; and
G is the product of a reaction between RCG1 and RCG2;
optionally in the presence of a modifying agent, to provide the compound of formula (V) above according to claim 1; and
(ii) optionally separating the compound of formula (V) provided in step (i) above from any unreacted compound of formula (IV) or unreacted antibody, Ab, or a combination thereof.