US 2013 0309192A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2013/0309192 A1 Yurkovetskiy et al. (43) Pub. Date: Nov. 21, 2013
(54) AURISTATIN COMPOUNDS AND filed on Jun. 24, 2011, provisional application No. CONUGATES THEREOF 61/513,234, filed on Jul. 29, 2011, provisional appli cation No. 61/566,935, filed on Dec. 5, 2011, provi (71) Applicant: Mersana Therapeutics, Inc., sional application No. 61/605,618, filed on Mar. 1, Cambridge, MA (US) 2012, provisional application No. 61/618,499, filed on Mar. 30, 2012. (72) Inventors: Aleksandr V. Yurkovetskiy, Littleton, MA (US); Mao Yin, Needham, MA Publication Classification (US); Timothy B. Lowinger, Carlisle, MA (US); Joshua D. Thomas, Natick, (51) Int. Cl. MA (US); Charles E. Hammond, A647/48 (2006.01) Billerica, MA (US); Cheri A. Stevenson, C07K 7/02 (2006.01) Haverhill, MA (US); Natalya D. (52) U.S. Cl. Bodyak, Brookline, MA (US); Patrick CPC ...... A6IK 47/48215 (2013.01); C07K 7/02 R. Conlon, Wakefield, MA (US); (2013.01) Dmitry R. Gumerov, Waltham, MA USPC ...... 424/78.17:530/330; 525/54.1: 514/21.8 (US) (57) ABSTRACT (73) Assignee: MERSANATHERAPEUTICS, INC., Auristatin compounds and conjugates thereof are provided Cambridge, MA (US) herein. The conjugate comprises a protein based recognition molecule (PBRM) and a polymeric carrier substituted with (21) Appl. No.: 13/944,561 one or more-LP-D, the protein based recognition-molecule being connected to the polymeric carrier by L. Each occur (22) Filed: Jul. 17, 2013 rence of D is independently an Auristatin compound. Land L are linkers connecting the therapeutic agent and PBRM to Related U.S. Application Data the polymeric carrier respectively. Also disclosed are poly meric scaffolds useful for conjugating with a PBRM to form (62) Division of application No. 13/493,899, filed on Jun. a polymer-drug-PBRM conjugate described herein, compo 11, 2012. sitions comprising the conjugates, methods of their prepara (60) Provisional application No. 61/495,771, filed on Jun. tion, and methods of treating various disorders with the con 10, 2011, provisional application No. 61/501,000, jugates or their compositions. Patent Application Publication Nov. 21, 2013 Sheet 1 of 8 US 2013/0309192 A1
700-.9F9in...... ty''''''''''''''''''''''''' 6 3. 7 -O Vehicle 600 -v- Example 8, 15.6 mg/kg 5500 -0- Example 8, 5.2 mg/kg 400 -e- Example 8, 1.6 mg/kg 300 W -E Example 8, 0.5 mg/kg -A Example 6 as 200 E 100 O HWE N to 3 - 9 S · S (S - MN 3 Days Post Dose
Figure 1 Patent Application Publication Nov. 21, 2013 Sheet 2 of 8 US 2013/0309192 A1
1000 ...seen......
6. Or Vehicle E 800 H Trastuzumab, 15 mg/kg S. -0- Example 7, 7.5 mg/kg 600 -- Example 6 + E Trastuzumab, 15 mg/kg is 400 -- Example 54, 20 mg/kg 5O 200
N & 9 - 9 $ ) Days Post Dose
Figure 2 Patent Application Publication Nov. 21, 2013 Sheet 3 of 8 US 2013/0309192 A1
1OOO ...F.R...... -...... -o- Vehicle 800 H Trastuzumab, 15 mg/kg -e- Example 51 600 +Trastuzumab, 15 mg/kg -E Example 52, 7.5 mg/kg 400
200
N 8 - 9 - S - Days Post Dose
Figure 3 Patent Application Publication Nov. 21, 2013 Sheet 4 of 8 US 2013/0309192 A1
Or Vehicle -V Example 7, 3.5 mg/kg dwkX3 -(e- Example 7, 10 mg/kg, qdx1 -- Example 7, 10 mg/kg, CWKX3
Y > Y S - Ky Y - 8 Yo S v So S v 8 (3) Days Post Dose
Figure 4 Patent Application Publication Nov. 21, 2013 Sheet 5 of 8 US 2013/0309192 A1
100 O Vehicle 468OOO ...Findpoint...... T.A...... - Example 60, 7 mg/kg 2 O
Y \ S S - Y - 8 Y so S v so S v S 3) Days Post Dose Figure 5 Patent Application Publication Nov. 21, 2013 Sheet 6 of 8 US 2013/0309192 A1
1,000,000
-0- Conjugated 3. 100,000 HPV S O S s 10,000 HPV Trastuzumab g 1OOO Conjugate 5 y (estimate) -O-Total 100 Trastuzumab L
1 O O 50 1OO 150 200
Time (hr)
Figure 6 Patent Application Publication Nov. 21, 2013 Sheet 7 of 8 US 2013/0309192 A1
9 OO
8OO
7OO
6OO
5OO
4OO
3OO
2OO 100
O 50 1 OO 150 2OO Time (hr)
Figure 7 Patent Application Publication Nov. 21, 2013 Sheet 8 of 8 US 2013/0309192 A1
1200 Vehicle
5 1000-...999...... T...... Example 70, 2 mg/kg Example 52, 2 mg/kg 8 O O Example 70, 4 mg/kg Example 52, 4 mg/kg 400
200
^ - V S K S 8 N 3° S) - so S 3 Days Post Dose
Figure 8 US 2013/0309192 A1 Nov. 21, 2013
AURISTATIN COMPOUNDS AND SUMMARY OF THE INVENTION CONUGATES THEREOF 0005. The present invention relates to a protein-polymer RELATED APPLICATIONS drug conjugate that is biodegradable, biocompatible and exhibits high drug load as well as strong binding to target 0001. This application is a division of U.S. patent appli antigen. The present invention also relates to a polymeric cation Ser. No. 13/493,899, filed Jun. 11, 2012, which claims scaffold useful to conjugate with a protein based recognition the benefit of and priority under 35 USC S119(e) to U.S. molecule (PBRM) so as to obtain the protein-polymer-drug Provisional Patent Application Nos. 61/495,771, filed Jun. conjugate. 10, 2011: 61/501,000, filed Jun. 24, 2011: 61/513,234, filed 0006. In one aspect, the invention features a polymeric Jul. 29, 2011: 61/566,935, filed Dec. 5, 2011: 61/605,618, scaffold useful to conjugate with a PBRM. The scaffold com filed Mar. 1, 2012; and 61/618,499, filed Mar. 30, 2012. The prises a polymeric carrier, one or more -L-D connected to contents of each of these applications are hereby incorporated the polymeric carrier, and one or more L connected to the by reference in their entireties. polymeric carrier which is suitable for connecting a PBRM to the polymeric carrier, wherein: BACKGROUND OF THE INVENTION 0007 each occurrence of D is independently a therapeutic 0002 Traditionally, pharmaceuticals have primarily con agent having a molecular weight s5 kDa, sisted of Small molecules that are dispensed orally (as Solid O008) the polymeric carrier is a polyacetal or polyketal, pills and liquids) or as injectables. Over the past three 0009 LP is a linker having the structure decades, formulations (i.e., compositions that control the route and/or rate of drug delivery and allow delivery of the therapeutic agent at the site where it is needed) have become increasingly common and complex. Nevertheless, many questions and challenges regarding the development of new treatments as well as the mechanisms with which to admin ister them remain to be addressed. For example, many drugs exhibit limited or otherwise reduced potencies and therapeu tic effects because they are either generally subject to partial with R' connected to an oxygen atom of the polymeric degradation before they reach a desired target in the body, or carrier and L' connected to D, and accumulate in tissues other than the target, or both. 0003. One objective in the field of drug delivery systems, therefore, is to deliver medications intact to specifically tar geted areas of the body through a system that can stabilize the drug and control the in vivo transfer of the therapeutic agent utilizing either physiological or chemical mechanisms, or both. denotes direct or indirect attachment of D to LP', and LP 0004 Antibody-drug conjugates have been developed as contains a biodegradable bond so that when the bond is bro target-specific therapeutic agents. Antibodies against various ken, D is released from the polymeric carrier in an active form cancer cell-surface antigens have been conjugated with dif for its intended therapeutic effect; ferent cytotoxic agents that inhibit various essential cellular targets such as microtubules (maytansinoids, auristatins, tax 10010 L' is a carbonyl-containing moiety; anes: U.S. Pat. Nos. 5,208,020; 5,416,064; 6,333.410; 6,441, 10011 L is a linker different from LP and having the struc 163; 6,340,701; 6,372,738; 6,436,931; 6,596,757; and 7,276, ture: —R' C(=O)-L' with R' connected to an oxygen 497); DNA (calicheamicin, doxorubicin, CC-1065 analogs; atom of the polymeric carrier and L' suitable for connecting U.S. Pat. Nos. 5,475,092: 5,585,499; 5,846,545; 6,534,660; directly or indirectly to a PBRM: 6.756,397; and 6,630.579). Antibody conjugates with some 0012 each of R'' and R' independently is absent, alkyl, of these cytotoxic drugs are actively being investigated in the heteroalkyl, cycloalkyl, or heterocycloalkyl, and clinic for cancer therapy (Ricart, A. D., and Tolcher, A. W., I0013 L' is a moiety containing a functional group that is 2007, Nature Clinical Practice, 4, 245-255; Krop et al., 2010, capable of forming a covalent bond with a functional group of J. Clin. Oncol., 28, 2698-2704). However, existing antibody a PBRM. drug conjugates have exhibited a few limitations. A major limitation is their inability to deliver a sufficient concentra 0014. The polymeric scaffold can include one or more of tion of drug to the target site because of the limited number of the following features. targeted antigens and the relatively moderate cytotoxicity of 10015 L is a linker having the structure: cancer drugs like methotrexate, daunorubicin, maytansi noids, taxanes, and Vincristine. One approach to achieving significant cytotoxicity is by linkage of a large number of drug molecules either directly or indirectly to the antibody. However such heavily modified antibodies often display impaired binding to the target antigen and fast in vivo clear ance from the blood stream. Therefore, there is a need to improve the ability to deliver a sufficient concentration of a in which L is a moiety containing a functional group that is drug to the target such that maximum cytotoxicity for the drug capable of forming a covalent bond with a functional group of is achieved. a PBRM, and US 2013/0309192 A1 Nov. 21, 2013
300), m is an integer from 1 to about 40, m is an integer from 1 to about 18, and/orm is an integer from 1 to about 140 (e.g., m being about 1-90). 0030. When the PHF in Formula (Ia) has a molecular weight ranging from about 6 kDa to about 20 kDa (i.e., the denotes direct or indirect attachment of L'' to L'. Sum of m, m, m, and m ranging from about 45 to about 0016. The functional group of L' or L is selected from 150), m is an integer from 2 to about 20, m is an integer from —SR, —S S-LG, maleimido, and halo, in which LG is a 1 to about 9, and/or m is an integer from 1 to about 75 (e.g., leaving group and R is H or a Sulfur protecting group. m being about 4-45). 0017 LP comprises —X-(CH), C(=O)— with X 0031 When the PHF in Formula (Ia) has a molecular directly connected to the carbonyl group of R' C(=O), in weight ranging from about 8 kDa to about 15 kDa (i.e., the which X is CH, O, or NH, and v is an integer from 1 to 6. sum of m, m, m, and m ranging from about 60 to about 0018. L' or L* contains a biodegradable bond. 110), m is an integer from 2 to about 15, m is an integer from 0019. Each of R'' and R is absent. 1 to about 7, and/or m is an integer from 1 to about 55 (e.g., 0020. The polymeric carrier of the scaffold of the inven m being about 4-30). tion is a polyacetal, e.g., a PHF having a molecular weight 0032. When the PHF in Formula (Ia) has a molecular (i.e., MW of the unmodified PHF) ranging from about 2 kDa weight ranging from 20 kDa to 300 kDa (i.e., the sum of m, to about 300 kDa. m, m, and m ranging from about 150 to about 2200), m is 0021 For conjugating a PBRM having a molecular weight an integer from 3 to about 300, m is an integer from 1 to about of 40 kDa or greater (e.g., 80 kDa or greater), the polymeric 110, and/orm is an integer from 1 to about 660 (e.g., m being carrier of the scaffold of the invention is a polyacetal, e.g., a PHF having a molecular weight (i.e., MW of the unmodified about 10-250). PHF) ranging from about 2 kDa to about 40 kDa (e.g., about 0033. When the PHF in Formula (Ia) has a molecular 6-20 kDa or about 8-15 kDa). weight ranging from 40 kDa to 150 kDa (i.e., the sum of m, 0022. For conjugating a PBRM having a molecular weight m, m, and m ranging from about 300 to about 1100), m is of 200 kDa or less (e.g., 80 kDa or less), the polymeric carrier an integer from 4 to about 150, m is an integer from 1 to about of the scaffold of the invention is a polyacetal, e.g., a PHF 75, and/orm is an integer from 1 to about 330 (e.g., m being having a molecular weight (i.e., MW of the unmodified PHF) about 15-100). ranging from about 20 kDa to about 300 kDa (e.g., about 0034. When the PHF in Formula (Ia) has a molecular 40-150 kDa or about 50-100 kDa). weight ranging from about 50 kDa to about 100 kDa (i.e., the 0023 The scaffold is of Formula (Ia): Sum of m, m, m, and m ranging from about 370 to about 740), m is an integer from 5 to about 100, m is an integer from 1 to about 40, and/orm is an integer from 1 to about 220 (Ia) (e.g., m being about 15-80). 0035. The scaffold further comprises a PBRM connected to the polymeric carrier via L. 0036. One or more PBRMs are connected to one drug carrying polymeric carrier. - 0037. The scaffold (e.g., a PBRM-polymer-drug conju hoc C L gate) is of Formula (Ib):
(Ib) O
3. Oo- in Oo- D iii. otho O D LP2 hocolL wherein: 0024 m is an integer from 1 to about 2200, 0025 m is an integer from 1 to about 660, O 0026 m is an integer from 1 to about 300, 0027 m is an integer from 1 to about 110, and 3 Oc D 0028 the sum of m, m, mand m ranges from about 15 o-J., L - J. to about 2200. 0029 When the PHF in Formula (Ia) has a molecular otho weight ranging from about 2 kDa to about 40 kDa (i.e., the D LP2 Sum of m, m, m, and m ranging from about 15 to about US 2013/0309192 A1 Nov. 21, 2013
-continued
O O
r OH O OH OH ={ LDI
OH O OH O O= O= in in wherein: LDI LDI
D LP2
OH O 0038 between Land PBRM denotes direct or indirect O ={ it 4 attachment of PBRM to L. LDI s 0039 each occurrence of PBRM independently has a molecular weight of less than 200 kDa, otho LP2 0040 m is an integer from 1 to about 2200, 0041 m is an integer from 1 to about 660, 0042 m is an integer from 3 to about 300, 0043 m is an integer from 0 to about 110, wherein: 0044 m is an integer from 1 to about 60; and 0050 terminal 0.045 the sum of m, m, m, m- and m ranges from about 150 to about 2200. 0046. In Formula (Ib), m is an integer from about 10 to about 660 (e.g., about 10-250). 0047. When the PHF in Formula (Ib) has a molecular weight ranging from 40 kDa to 150 kDa (i.e., the sum of m, attached to L denotes director indirect attachment of L° to m, m, m, and m ranging from about 300 to about 1100), PBRM such that the D-carrying polymeric carrier is con m is an integer from 4 to about 150, m is an integer from 1 nected to the PBRM, to about 75, m is an integer from 1 to about 30, and/or m is 0051 m is an integer from 1 to 300, an integer from 1 to about 330 (e.g., m being about 10-330 or 0.052 m is an integer from 1 to 140, about 15-100). 0053 m is an integer from 1 to 40, 0048. When the PHF in Formula (Ib) has a molecular 0054 m is an integer from 0 to 18, weight ranging from about 50 kDa to about 100 kDa (i.e., the 0055 m is an integer from 1 to 10; and Sum of m, m, m2, m, and m ranging from about 370 to 0056 the sum of m, m, m ms, and m ranges from 15 to about 740), m is an integer from 5 to about 100, m is an 300; provided that the total number of L'attached to the integer from 1 to about 40, m is an integer from 1 to about 20, PBRM is 10 or less. and/or m is an integer from 1 to about 220 (e.g., m being 0057. In Formula (Ic), m is an integer from 1 to about 120 about 15-80). (e.g., about 1-90) and/or m is an integer from 1 to about 10 0049. Alternatively or additionally, one or more drug-car (e.g., about 1-8). rying polymeric carriers are connected to one PBRM. The 0058 When the PHF in Formula (Ic) has a molecular scaffold (e.g., a PBRM-polymer-drug conjugate) comprises a weight ranging from about 6 kDa to about 20 kDa (i.e., the PBRM with a molecular weight of greater than 40 kDa and Sum of m, m1, m2, m3, and m ranging from about 45 to about one or more D-carrying polymeric carriers connected to the 150), m is an integer from 2 to about 20, m is an integer from PBRM, in which each of the D-carrying polymeric carrier 1 to about 9, and/or m is an integer from 1 to about 75 (e.g., independently is of Formula (Ic): m being about 4-45). US 2013/0309192 A1 Nov. 21, 2013
0059. When the PHF in Formula (Ic) has a molecular when not connected to PBRM, independently comprises a weight ranging from about 8 kDa to about 15 kDa (i.e., the terminal group W, in which each W independently is: Sum of m, m. m. m., and m ranging from about 60 to about 110), m is an integer from 2 to about 15, m is an integer from 1 to about 7, and/or m is an integer from 1 to about 55 (e.g., (1) m being about 4-30). 0060 Each occurrence of D independently is selected from Vinca alkaloids, auristatins, tubulysins, duocarmycins, kinase inhibitors, MEK inhibitors, KSP inhibitors, and ana (2) logs thereof. 0061 LP is R. C(O) X-MP-Y-MP-Z'- MP-QP-MP- with MP directly connected to D, in which (3) I0062 XP is —O , —S , N(R') , or absent, in which R' is hydrogen, an aliphatic, heteroaliphatic, carbocy clic, or heterocycloalkyl moiety, —C(=O)R', —C(=O) OR', or—SOR', or N(R') is a heterocycloalkyl moi (4) ety, wherein R' is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety;
(5)
(6)
(7)
O(NR) , -(CR—N)O , C(=O)NR (N=CR) , -(CR—N) NRC(=O) , -SO, , (8) -NRSONR , -SONR , and polyamide, wherein each occurrence of R. R. and Rindependently is hydrogen or an aliphatic, heteroaliphatic, carbocyclic, or heterocyclic moiety, or each occurrence of NR or NRNR is a (9) heterocycloalkyl moiety; and 0064 each of M', MP, MP, and Mindependently, is absent or a non-biodegradable linker moiety selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, het eroalkenyl, heteroalkynyl, a carbocyclic moiety, a heterocy clic moiety, and a combination thereof, and each of M', (10) M', and M' optionally contains one or more —(C=O)— but does not contain any said biodegradable linker moiety; 0065 provided that for each L', at least one of X, Y, ZP, and QP is not absent. 0066. Each (11)
LD LP2 US 2013/0309192 A1 Nov. 21, 2013
-continued in which R' is a leaving group (e.g., halide or RC(O)O— in (12) which R is hydrogen, an aliphatic, heteroaliphatic, carbocy clic, or heterocycloalkyl moiety), R'' is a sulfur protecting N SN1 group, and ring A is cycloalkylorheterocycloalkyl, and R' is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or het erocycloalkyl moiety. (13) 0067. Each R' independently is
O
RSI RS2 All - Pl (14) FCOORs.3 rt Rs,
O RSI RS2 Rs COORs.3 YOSORS: RS2 (15) Rs 1 COORs.3 HO in which ris 1 or 2 and each of R', R, and R is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl (16) moiety. 0068. Each
(17) LD LP2
when connected to PBRM, independently is X-M'-Y- M°-Z-M-Q-M-, with X directly connected to the carbonyl group of R'—C(=O) and M. directly connected (18) to PBRM, in which 0069 X is -O-, -S-, - N(R') , or absent, in which R" is hydrogen, an aliphatic, heteroaliphatic, carbocy clic, or heterocycloalkyl moiety, —C(=O)R', C(=O) OR', or -SOR', or N(R') is a heterocycloalkyl moi (19) ety, wherein R' is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety; (0070 each of Y, Z, and Q, independently, is absent or a biodegradable linker moiety selected from the group con sisting of S S , —C(=O)C) , —C(=O)NR' ,
(20)
(21)
(22) O(N–CR) , (CR-N)O C(=O)NR (NR) , -(CR=N) NRC(=O) , —SO , - NRSONR , -SONR , and polyamide, wherein each occurrence of R. R. and Rindependently is hydrogen US 2013/0309192 A1 Nov. 21, 2013
or an aliphatic, heteroaliphatic, carbocyclic, or heterocyclic -continued moiety, or each occurrence of NR— or NRNR is a O S heterocycloalkyl moiety; and p
(0071 each of M', M', M, and Mindependently, is N s absent or a non-biodegradable linker moiety selected from the (CH3)1-2 group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, het eroalkenyl, heteroalkynyl, a carbocyclic moiety, a heterocy O clic moiety, and a combination thereof, and each of M', M. and M optionally contains one or more —(C=O)—but does not contain any said biodegradable linker moiety; 0072 provided that for each
o LD LP2 connected to PBRM, at least one of X, Y, Z, and Q is not absent. Ns 0073. Each of MP' and M' independently is Calkyl or C X. C. heteroalkyl. 0074 Each of MP, MP, MP, M2, M, and M, inde pendently is absent, Ce alkyl, cycloalkyl, heteroalkyl, het and erocycloalkyl, or a combination thereof. 21 0075. In each -- N
LD LP2 at most one of M and M has one of the following struc tures: in which q is an integer from 0 to 12 and each of p and t independently is an integer from 0 to 3. 0076 Also within the scope of the invention is a method of preparing a scaffold described above. The method comprises providing a polymeric carrier that is Substituted with one or more-LP-Dandone or more R' C(=O)-L', and react ing the polymeric carrier with a compound containing an L. moiety to produce the scaffold of claim 2 comprising a poly meric carrier substituted both with one or more-LP-D and with one or more
0077 Alternatively, the method comprises providing a polymeric carrier that is substituted with one or more
and one or more —R' C(=O)-LP', and reacting the poly meric carrier with D containing a functional group that is US 2013/0309192 A1 Nov. 21, 2013 capable of forming a covalent bond with —R'' C(=O)- -continued LP to produce the scaffold of claim 2 comprising a polymeric carrier substituted both with one or more-LP-D and with one OO
(6) O
-R1-C(=O)-LP LP2. A- O --- CH3 0078. The invention also features a compound of Formula (XII) or (XIIa):
(XII) H3C CH3 H3C O CH3 CH3
HC3 NN Null N O
CH, O E CH, OCH O OCH O H3C 1Net, O N-R (XIIa) HC CH HC O CH CH HC N 3 NN N CH, O CH, OCH O OCH O HC CH O N-R H or a pharmaceutically acceptable salt thereof, -continued wherein O (7)
22 0079 Rao is selected from the group consisting of x-r O lunt
(1) (8) O OH: NH2: x-r O (2) OH: CH, O (9)
CH O --- NH2: (3) OH: (10) CH, O
(4) O --- NH2: OH: (11) * -- O CH (5) (12) NH2: CH, O CH3
>n-> O CH -sul US 2013/0309192 A1 Nov. 21, 2013
-continued nition-molecule (PBRM) and a therapeutic agent (D). The (13) scaffold (i.e., the one free of any D) comprises a polymeric O CH3 carrier, one or more L connected to the polymeric carrier which is suitable for connecting a PBRM to the polymeric O --- NH2: carrier, and one or more —R'' C(=O)-L' connected to (14) the polymeric carrier via R', wherein: CH, O I0084 the polymeric carrier is a polyacetal or polyketal, NH2: I0085) R' is connected to an oxygenatom of the polymeric O carr1er, I0086) LP is a linker suitable for connecting a D molecule CH to the polymeric carrier, in which each occurrence of D is (15) independently a therapeutic agent having a molecular weight CH, O s5 kDa; I0087 L is a linker different from -R-C(=O)-LP', O and having the structure:-R' C(=O)-L" with R' con nected to an oxygen atom of the polymeric carrier and L' (16) suitable for connecting to a PBRM; O I0088 each of R'' and R' independently is absent, alkyl, heteroalkyl, cycloalkyl, or heterocycloalkyl; I0089) L' is a moiety containing a functional group that is >is ---1-2 N-) NH3. capable of forming a covalent bond with a functional group of D, and (17) I0090. L' is a moiety containing a functional group that is capable of forming a covalent bond with a functional group of a PBRM. 0091. The D-free scaffold useful to conjugate with a (18) PBRM and a D can have one or more of the following fea tures. 0092 L is a linker having the structure: (19) O H N x-rayO and (20) O CH3 in which L is a moiety containing a functional group that is capable of forming a covalent bond with a functional group of > --> 1. a PBRM, and NH2
0080 a is an integer from 1 to 6; and 0081 c is an integer from 0 to 3. 0082 Rao can be denotes direct or indirect attachment of L to L'. (0093. The functional group of L' or L is selected from —SRP. —S S-LG, maleimido, and halo, in which LG is a leaving group and RP is H or a sulfur protecting group. > --> 0094 LP' comprises —X-(CH), C(=O)— with X O directly connected to the carbonyl group of R'' C(=O), in which X is CH, O, or NH, and V is an integer from 1 to 6. X-----O (0095 L or L* contains a biodegradable bond. CH 0096. Each of R'' and R is absent. (0097. The polymeric carrier of the D-free scaffold is a polyacetal, e.g., a PHF having a molecular weight (i.e., MW of the unmodified PHF) ranging from about 2 kDa to about CH3 300 kDa. 0098. For conjugating a PBRM having a molecular weight 0083. In another aspect, the invention features a polymeric of 40 kDa or greater (e.g., 80 kDa or greater), the polymeric scaffold useful to conjugate with both a protein based recog carrier of the D-free scaffold is a polyacetal, e.g., a PHF US 2013/0309192 A1 Nov. 21, 2013
having a molecular weight (i.e., MW of the unmodified PHF) 0110. When the PHF in Formula (Id) has a molecular ranging from about 2 kDa to about 40 kDa (e.g., about 6-20 weight ranging from about 50 kDa to about 100 kDa (i.e., the kDa or about 8-15 kDa). Sum of m, m, and m ranging from about 370 to about 740), 0099 For conjugating a PBRM having a molecular weight m is an integer from 1 to about 40, and/or m is an integer of 200 kDa or less (e.g., 80 kDa or less), the polymeric carrier from 1 to about 220 (e.g., m being about 20-180). of the D-free scaffold of the invention is a polyacetal, e.g., a 0111. The D-free scaffold further comprises a PBRM con PHF having a molecular weight (i.e., MW of the unmodified nected to the polymeric carrier via L. PHF) ranging from about 20 kDa to about 300 kDa (e.g., 0112. One or more PBRMs are connected to one D-free about 40-150 kDa or about 50-100 kDa). polymeric carrier. 0100. The D-free scaffold is of Formula (Id): 0113. The D-free scaffold is of Formula (Ie):
(Id) (Ie) O O r OH O OH OH OH OH iii. o={ in iii. LDI LDI
OH O OH O
o={ it 3. O ={ in 3 LDI LDI
LP2 LP2 wherein: 0101 m is an integer from 1 to about 2200, 0102 m is an integer from 1 to about 660, OH O (0103 m is an integer from 1 to about 110, and O ={ it 4 0104 the sum of m, m, and m ranges from about 15 to LDI about 2200. 0105. When the PHF in Formula (Id) has a molecular weight ranging from about 2 kDa to about 40 kDa (i.e., the sum of m, m, and m ranging from about 15 to about 300), m LP2 is an integer from 1 to about 18, and/or m is an integer from 1 to about 140 (e.g., m being about 2-120). PBRM, 0106. When the PHF in Formula (Id) has a molecular weight ranging from about 6 kDa to about 20 kDa (i.e., the Sum of m, m, and m ranging from about 45 to about 150), m wherein: is an integer from 1 to about 9, and/orm is an integer from 1 to about 75 (e.g., m being about 6-60). 0107. When the PHF in Formula (Id) has a molecular weight ranging from about 8 kDa to about 15 kDa (i.e., the Sum of m, m, and m ranging from about 60 to about 110), m is an integer from 1 to about 7, and/orm is an integer from 1 to about 55 (e.g., m being about 6-45). 0108. When the PHF in Formula (Id) has a molecular 0114 between Land PBRM denotes direct or indirect weight ranging from 20 kDa to 300 kDa (i.e., the sum of m, attachment of PBRM to L’, m, and m ranging from about 150 to about 2200), m is an 0115 PBRM has a molecular weight of less than 200 kDa, integer from 1 to about 110, and/orm is an integer from 1 to 0116 m is an integer from 1 to 2200, about 660 (e.g., m being about 13-550). 0117 m is an integer from 1 to 660, 0109. When the PHF in Formula (Id) has a molecular weight ranging from 40 kDa to 150 kDa (i.e., the sum of m, I0118 m is an integer from 0 to 110, m, and m ranging from about 300 to about 1100), m is an I0119 m is an integer from 1 to about 60; and integer from 1 to about 75, and/or m is an integer from 1 to 0120 the sum of m, m, m, m- and m ranges from about about 330 (e.g., m being about 20-250). 150 to about 2200. US 2013/0309192 A1 Nov. 21, 2013
0121. In Formula (Ie), m is an integer from about 10 to 0.126 m is an integer from 1 to 300, about 660 (e.g., about 14-550). I0127 m is an integer from 1 to 140, 0122) When the PHF in Formula (Ie) has a molecular I0128 m is an integer from 0 to 18, weight ranging from 40 kDa to 150 kDa (i.e., the sum of m, I0129 m is an integer from 1 to 10; and m, m, and m ranging from about 300 to about 1100), m is 0.130 the sum of m, m, m, and m ranges from 15 to 300; an integer from 1 to about 75, ma is an integer from 1 to about provided that the total number of Lattached to the PBRM is 30, and/orm is an integer from 1 to about 330 (e.g., m being 10 or less about 20-250). I0131. In Formula (Ih), m is an integer from 2 to about 130 (0123. When the PHF in Formula (Ie) has a molecular (e.g., about 3-120) and/or m is an integer from 1 to about 10 weight ranging from about 50 kDa to about 100 kDa (i.e., the (e.g., about 1-8). sum of m, m, m, and m ranging from about 370 to about (0132) When the PHF in Formula (Ih) has a molecular 740), m is an integer from 1 to about 40, m is an integer from weight ranging from about 6 kDa to about 20 kDa (i.e., the 1 to about 20, and/orm is an integer from 1 to about 220 (e.g., Sum of m, m, m, and m ranging from about 45 to about m being about 20-180). 150), m is an integer from 1 to about 9, and/orm is an integer 0.124. Alternatively or additionally, one or more D-free from 6 to about 75 (e.g., m being about 7-60). polymeric carriers are connected to one PBRM. The scaffold I0133. When the PHF in Formula (Ih) has a molecular comprises a PBRM with a molecular weight of greater than weight ranging from about 8 kDa to about 15 kDa (i.e., the 40 kDa and one or more polymeric carriers connected to the Sum of m, m, m, and m ranging from about 60 to about PBRM, in which each of the polymeric carrier independently 110), m is an integer from 1 to about 7, and/orm is an integer is of Formula (Ih): from 6 to about 55 (e.g., m being about 7-45). I0134. As used herein, the terms “polymeric scaffold' or simply “scaffold' and "conjugate' are used interchangeably (Ih) when the scaffold comprises one or more PBRM and one or more D molecules. I0135) In yet another aspect, the invention encompasses a conjugate comprising a polymeric carrier, one or more-LP-D connected to the polymeric carrier, and a protein based rec cC OH O ognition-molecule (PBRM) connected to the polymeric car m rier via L., wherein: o={LDI 0.136 each occurrence of D is independently a therapeutic agent (e.g., a drug) having a molecular weightss kDa, 0.137 the polymeric carrier is a polyacetal or polyketal, 0138 LP is a linker having the structure: - R - C (-O) XP-MP1-YP-MP2-ZP-MP-QD-MP4-, with R?.1 connected to an oxygen atom of the polymeric carrier and MP connected to D; C 0.139 L is a linker having the structure: - R' C (—O) X-M-Y-MP-Z-M.-Q-M-, with R.? con nected to an oxygen atom of the polymeric carrier and M connected to the protein based recognition-molecule: I0140 each of R'' and R' independently is absent, alkyl, cycloalkyl, heteroalkyl, or heterocycloalkyl: I0141 each of XP and X, independently is—O ,—S , —N(R')—, or absent, in which R' is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety, C(=O)R', C(=O)CR', SOR' or N(R') is a C heterocycloalkyl moiety, wherein R' is hydrogen, an ali phatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moi ety; 0.142 each ofYP,Y,Z, Z, QP, and Q, independently, is absent or a biodegradable linker moiety selected from the group consisting of —S—S , —C(=O)C)— —C(=O) wherein: NR OC(=O) NRC(=O) OC(=O)C 0125 terminal OC(=O)NR NRC(=O)C NRC(=O) NR C(OR)O C(OR)S C(OR)NR
attached to L denotes director indirect attachment of L to PBRM such that the D-carrying polymeric carrier is con nected to the PBRM, US 2013/0309192 A1 Nov. 21, 2013 11
NRNRC(=O)C , C(-S)NRNR NRNRC -continued ( S) , C(-NR)NRNR , NRNRC(-NR) , (7) O(N–CR) , (CR-N)O C(=O)NR O (NR) , (CR-N) NRC(=O) , —SO , -NRSONR , -SONR , and polyamide, wherein NHNH2: each occurrence of R. R. and Rindependently is hydrogen or an aliphatic, heteroaliphatic, carbocyclic, or heterocyclic (8) moiety, or each occurrence of NR or NRNR is a heterocycloalkyl moiety; and 0143 each of MP, MP, MP, MP, M', MP, M and O M", independently, is absent or a non-biodegradable linker (9) moiety selected from the group consisting of alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, a car bocyclic moiety, a heterocyclic moiety, and a combination thereof, and each of MP, MP, MP, M', M, and M optionally contains one or more —(C=O)— but does not Rl'. contain any said biodegradable linker moiety; (10) 0144) provided that for each LP, at least one of XP, YP. ZP, and QP is not absent, and for each L. at least one of X, Y, Z, and Q is not absent. OMe: 0145 The conjugate can include one or more of the fol- -Ph lowing features. 0146 The polymeric carrier can be a polyacetal, e.g., PHF. Ph 0147 For each LP. M'P' is not absent when XP is absent. (11) 0148. For each L. M' is not absent when X is absent. N SC, 014.9 The polymeric carrier can be further substituted N with one or more R. C(=O) XP-MP-Y-MP-W), 2 in which each WP independently is: (12)
(1) N ON SH: (13) O; a x N SR14: O (3) (14)
(4)
NH2: o (15) (5)5 O
(16)
(6) (17) US 2013/0309192 A1 Nov. 21, 2013 12
-continued -continued (18) (29) Rld I OH: N
D s (19)
(30) (20) r y O (21) (31) N SX 2N O (22) (32) 21 Ns1 y O / \, (33) (23) -COOH
in which R'' is a sulfur protecting group, each of ring A and B, independently, is cycloalkyl or heterocycloalkyl, R' is an aliphatic, heteroaliphatic, carbocyclic or heterocycloalkyl (24) moiety; ring D is heterocycloalkyl, R' is hydrogen, an ali phatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moi ety; and R' is a leaving group (e.g., halide or RC(O)C)— in which R is hydrogen, an aliphatic, heteroaliphatic, carbocy clic, or heterocycloalkyl moiety). (25) 0150. The polymeric carrier can be further substituted with one or more R. C(=O) X-M-Y-MW, in which each Windependently is:
(1) (26) SH:
(2)
(27) SR14:
(3)
(28) (4) US 2013/0309192 A1 Nov. 21, 2013 13
-continued -continued (5) (15)
HO
(16) A.O Ac; (6)
NH (17) -- Rl. (7) O
X's. (18) (8)
o1 (9) SH (19)
(10) O (20)
OMe: r y O Ph (21) r O Ph S (11) N X s 2N O N SC, (22)
(12) N SN1n S s l in which R' is a leaving group (e.g., halide or RC(O)O— in ON which R is hydrogen, an aliphatic, heteroaliphatic, carbocy (13) clic, or heterocycloalkyl moiety), R'' is a sulfur protecting group, and ring A is cycloalkylorheterocycloalkyl, and R' is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or het 7 erocycloalkyl moiety. For example, R' is O (14) US 2013/0309192 A1 Nov. 21, 2013
-continued -continued Rs 1 COORs.3 RSI Rs2 Rs2 OSOR, Rs COOR3, in which ris 1 or 2 and each of R', R, and R is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety. 0151 Ring. A can be Cs cycloalkyl or 5-19 membered heterocycloalkyl. 0152 Ring. A can be A-C) C. ^ 0153 Ring B can be Cs cycloalkyl or 3-12 membered heterocycloalkyl. 0154 Ring D can be piperazinyl or piperidinyl. O155 Each of R', R, and R can be hydrogen or C. alkyl. 0156 Each PBRMindependently can be a peptide, a pep tide mimetic, an antibody, or an antibody fragment. 0157 Each of MP' and M' independently can be C. alkyl or C. heteroalkyl. 0158 Each of MP, MP, MP, M2, M, and M, inde pendently can be absent, Ce alkyl, cycloalkyl, heteroalkyl, heterocycloalkyl, or a combination thereof. 0159 For each LP, at most two of MP, MP, and MP can be absent. (0160 For each Lif, at most two of M, M, and M can be absent. (0161 For each LP, at most one of MP and MP can have one of the following structures:
in which q is an integer from 0 to 12 and each of p and t independently is an integer from 0 to 3. (0162 For each L. at most one of M* and M can have one of the following structures:
O -
xO -r US 2013/0309192 A1 Nov. 21, 2013 15
-continued (0163 For each LP, each of -MP-ZP ZP-MP-, —Z'-M-, and-Ms-Z, , independently can have one of
O th-i-q Y it s the following structures:
(1)
O O PYs s O (2) N f N O s O >> S SX N2N. (3) (CH3)1-2 l 7 s O S chp N-- (4) (CH3)1-2
O NH s
S it: ; x^121
21 p ry i" (6)6 S-1 s -At-A- (7)
O x's,NHN 21 pns ul- and \ s N O (8) CH3 N M O A / g s (9) S in which q is an integer from 0 to 12 and each of p and t x Rld independently is an integer from 0 to 3. US 2013/0309192 A1 Nov. 21, 2013 16
-continued -continued (10) (18)
(11) (19)
(12) (20)
(21) O (13) v-y O O NY (22) C “y f (14) O O As-SX.O O N O (23)23 ry - and O
Rld O N N - B (16) O O Y (24)
(17) O NHN in which ring A or B independently is cycloalkyl or hetero x cycloalkyl; R" is analiphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety; R' is hydrogen, an aliphatic, het eroaliphatic, carbocyclic, or heterocycloalkyl moiety; and ring D is heterocycloalkyl. US 2013/0309192 A1 Nov. 21, 2013 17
(0164. For each L', each of -M-Z - Zif-M-, Zif -continued M°-, and -M-Z , independently, can have one of the (10) following structures:
(1)
(11) (2)
(12) (3)
(4) in which ring A is cycloalkyl or heterocycloalkyl and R' is NH hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or het erocycloalkyl moiety. (0165. Each of XP and X, independently can be absent. (0166 Each of XP and X, independently can be O or NH. HO (0167. Each of XP and X, independently can be (5)
O OEC
N\ /N O o (6) (0168 Each ofYP and Y independently can be-S-S , OCO-, -COO – CONH , or - NHCO-. (0169. Each of Q' and Q independently can be absent, S–S OCO ,-COO - CONH NHCO , OCONHNH Or NHNHCOO . (7) 0170 In particular, this invention features a conjugate of Formula (I):
(I) (8) foci C. (9) US 2013/0309192 A1 Nov. 21, 2013 18
-continued 0.174. The ratio between n and n can be about 50:1, 40:1. 25:1, 20:1, 10:1, 5:1 or 2:1. 0.175. In another aspect, the invention provides composi tions comprising the conjugates, methods for their prepara OH O tion, and methods of use thereof in the treatment of various disorders, including, but not limited to cancer. O ={ 0176 The invention also features a drug-polymer conju "J,D gate (e.g., therapeutic agent-polymer conjugate) that is simi MPI lar to the protein-polymer-drug conjugate described above a YD except that drug-polymer conjugate does not contain a PBRM. In this embodiment the polymer-drug conjugate may MP2 comprise a plurality of drug moieties in which each D can be the same or different. In this embodiment, n is 0 in the D3 conjugate of Formula (I). The methods of producing the drug MS polymer conjugates and methods of treating various disorders MP4 (e.g., cancer) are also contemplated and described herein. ND 0177. The invention also features a protein-polymer con jugate (e.g., PBRM-polymer conjugate) that is similar to the protein-polymer-drug conjugate described above except that protein-polymer conjugate does not contain a drug. In this OH O embodiment the protein-polymer conjugate may comprise a plurality of protein moieties in which each PBRM can be the O ={ same or different. In this embodiment, n is 0 in the conjugate XP of Formula (I). The methods of producing the drug-polymer -n, MPI conjugates or polymeric scaffolds and methods of treating n YP various disorders (e.g., cancer) are also contemplated and described herein. The target cancer can be anal, astrocytoma, MP2 leukemia, lymphoma, head and neck, liver, testicular, cervi a. WP cal, sarcoma, hemangioma, esophageal, eye, laryngeal, mouth, mesothelioma, skin, myeloma, oral, rectal, throat, bladder, breast, uterus, ovary, prostate, lung, colon, pancreas, renal, or gastric cancer. OH cO 0.178 The invention further relates to a pharmaceutical O ={ composition comprising a polymeric scaffold or conjugate XP described herein and a pharmaceutically acceptable carrier. -n. 0179. In yet another aspect, the invention relates to a MPI n YP method of diagnosing a disorder in a subject Suspected of V having the disorder. The method comprises administering an MP2 A effective amount of the conjugate described herein to the ZP Subject Suspected of having the disorder or performing an V assay to detect a target antigen/receptor in a sample from the MP3 Subject so as to determine whether the Subject expresses target antigen or receptor. 0180. Unless otherwise defined, all technical and scien MP4 tific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this PBRM, invention belongs. In the specification, the singular forms also include the plural unless the context clearly dictates wherein each of n, n, n, n, and n is the molar fraction of otherwise. Although methods and materials similar or equiva the corresponding polymer unit ranging between 0 and 1: lent to those described herein can be used in the practice or n+n+n+n+n 1; provided that none of n, n, and n is 0. testing of the present invention, Suitable methods and mate 0171 In Formula (I) above, the disconnection or gap rials are described below. All publications, patent applica between the polyacetal units indicates that the units can be tions, patents and other references mentioned herein are connected to each other in any order. In other words, the incorporated by reference. The references cited herein are not appending groups that contain D, PBRM, W, and W, can be admitted to be prior art to the claimed invention. In the case of randomly distributed along the polymer backbone. conflict, the present specification, including definitions, will 0172. In the protein-polymer-drug conjugate of Formula control. In addition, the materials, methods and examples are (I), each D can be the same or different moiety and each illustrative only and are not intended to be limiting. PBRM can be the same or different moiety. 0181. One of the advantages of the present invention is that 0173 The ratio between n and n can be greater than 1:1, the protein-polymer-drug conjugates or the polymeric scaf and up to 200:1 (e.g., up to 100:1), e.g., between 2:1 and 40:1; folds described herein greatly enhances the bioavailability of between 5:1 and 20:1; between 10:1 and 50:1, between 25:1 the drugs to be delivered and/or enhances the bioavailability and 50:1, or between 30:1 and 50:1. of the protein attached to the polymeric carrier. Other features
US 2013/0309192 A1 Nov. 21, 2013 20 can be more narrowly claimed using the intermediate term groups include, but are not limited to halides such as Cl, Br, “consisting essentially of or the closed term “consisting of.” and I, Sulfonate esters, such as para-toluenesulfonate (“tosy 0.195 Recitation of ranges of values are merely intended to late’, TsO), and RC(O)C)— in which R is hydrogen, an serve as a shorthand method of referring individually to each aliphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl separate value falling within the range, unless otherwise indi moiety. cated herein, and each separate value is incorporated into the 0198 All methods described herein can be performed in specification as if it were individually recited herein. A range any suitable order unless otherwise indicated herein or oth used herein, unless otherwise specified, includes the two lim erwise clearly contradicted by context. The use of any and all its of the range. For example, the expressions "X being an examples, or exemplary language (e.g., “such as') provided integer between 1 and 6” and “x being an integer of 1 to 6’ herein, is intended merely to better illustrate the invention and both mean "x being 1, 2, 3, 4, 5, or 6”. is not to be construed as a limitation on the scope of the claims 0196) “Protecting group': as used herein, the term protect unless explicitly otherwise claimed. No language in the speci ing group means that a particular functional moiety, e.g., O.S, fication is to be construed as indicating that any non-claimed or N, is temporarily blocked so that a reaction can be carried element is essential to what is claimed. out selectively at another reactive site in a multifunctional 0199 Antibody” refers to an immunoglobulin molecule compound. In preferred embodiments, a protecting group of the class IgG including but not limited to IgG subclasses reacts selectively in good yield to give a protected Substrate (IgG1, 2, 3 and 4) and class IgM which is able to specifically that is stable to the projected reactions; the protecting group bind to a specific epitope on an antigen. Antibodies can be must be selectively removed in good yield by readily avail intact immunoglobulins derived from natural sources or from able, preferably nontoxic reagents that do not attack the other recombinant sources and can be immunoreactive portions of functional groups; the protecting group forms an easily sepa intact immunoglobulins. Antibodies may exist in a variety of rable derivative (more preferably without the generation of forms including, for example, polyclonal antibodies, mono new stereogenic centers); and the protecting group has a clonal antibodies, camelized single domain antibodies, intra minimum of additional functionality to avoid further sites of cellular antibodies (“intrabodies'), recombinant antibodies, reaction. As detailed herein, oxygen, Sulfur, nitrogen and anti-idiotypic antibodies, domain antibodies, linear antibody, carbon protecting groups may be utilized. For example, in multispecific antibody, antibody fragments, such as, Fv, Fab, certain embodiments, certain exemplary oxygen protecting Fab', Fab'-SH, F(ab'), single chain variable fragment anti groups may be utilized. These oxygen protecting groups bodies (scFv), Fc, pFc', scFvFc, disulfide Fv (dsfv), bispecific include, but are not limited to methyl ethers, substituted antibodies (bc-scFv) such as BiTE antibodies; camelid anti methyl ethers (e.g., MOM (methoxymethyl ether), MTM bodies, resurfaced antibodies, humanized antibodies, fully (methylthiomethyl ether), BOM (benzyloxymethyl ether), human antibodies, single-domain antibody (SdAb, also and PMBM (p-methoxybenzyloxymethyl ether)), substituted known as NANOBODYR), chimeric antibodies, chimeric ethyl ethers, substituted benzyl ethers, silyl ethers (e.g., TMS antibodies comprising at least one human constant region, (trimethylsilyl ether), TES (triethylsilylether), TIPS (triso dual-affinity antibodies such as, dual-affinity retargeting pro propylsilyl ether), TBDMS (t-butyldimethylsilyl ether), teins (DARTTM), divalent (or bivalent) single-chain variable tribenzylsilyl ether, and TBDPS (t-butyldiphenylsilyl ether), fragments (di-scFVs, bi-sch vs) including but not limited to esters (e.g., formate, acetate, benzoate (BZ), trifluoroacetate, minibodies, diabodies, triabodies or tribodies, tetrabodies, and dichloroacetate), carbonates, cyclic acetals and ketals. In and the like, and multivalent antibodies. Antibody fragment' certain other exemplary embodiments, nitrogen protecting refers to at least a portion of the variable region of the immu groups are utilized. Nitrogen protecting groups, as well as noglobulin molecule that binds to its target, i.e., the antigen protection and deprotection methods are known in the art. binding region. As used herein, the term “antibody” refers to Nitrogen protecting groups include, but are not limited to, both the full-length antibody and antibody fragments unless carbamates (including methyl, ethyl and Substituted ethyl otherwise specified. carbamates (e.g., Troc), amides, cyclic imide derivatives, (0200) “Protein based recognition-molecule' or “PBRM’ N-Alkyl and N-Aryl amines, imine derivatives, and enamine refers to a molecule that recognizes and binds to a cell Surface derivatives. In yet other embodiments, certain exemplary Sul marker or receptor Such as, a transmembrane protein, Surface phur protecting groups may be utilized. The Sulfur protecting immobilized protein, or protoglycan. Examples of PBRMs groups include, but are not limited to those oxygen protecting include but are not limited to, antibodies (e.g., Trastuzumab, group describe above as well as aliphatic carboxylic acid Cetuximab, Rituximab, Bevacizumab, Epratuzumab, Veltu (e.g., acrylic acid), maleimide, vinyl Sulfonyl, and optionally Zumab, LabetuZumab) or peptides (LHRH receptor targeting Substituted maleic acid. Certain other exemplary protecting peptides, EC-1 peptide), lipocalins, such as, for example, groups are detailed herein, however, it will be appreciated that anticalins, proteins such as, for example, interferons, lym the present invention is not intended to be limited to these phokines, growth factors, colony stimulating factors, and the protecting groups; rather, a variety of additional equivalent like, peptides or peptide mimics, and the like. The protein protecting groups can be readily identified using the above based recognition molecule, in addition to targeting the modi criteria and utilized in the present invention. Additionally, a fied polymer conjugate to a specific cell, tissue or location, variety of protecting groups are described in “Protective may also have certain therapeutic effect such as antiprolifera Groups in Organic Synthesis’ Third Ed. Greene, T. W. and tive (cytostatic and/or cytotoxic) activity against a target cell Wuts, P.G., Eds. John Wiley & Sons, New York: 1999, the or pathway. The protein based recognition molecule com entire contents of which are hereby incorporated by refer prises or may be engineered to comprise at least one chemi CCC. cally reactive group Such as, -COOH, primary amine, sec 0197) “Leaving group' refers to a molecular fragment that ondary amine —NHR, SH, or a chemically reactive amino departs with a pair of electrons in heterolytic bond cleavage. acid moiety or side chains such as, for example, tyrosine, Leaving groups can be anions or neutral molecules. Leaving histidine, cysteine, or lysine. US 2013/0309192 A1 Nov. 21, 2013
0201 “Biocompatible' as used herein is intended to exclusion HPLC. Although faster degradation may be in describe compounds that exert minimal destructive or host Some cases preferable, in general it may be more desirable response effects while in contact with body fluids or living that the polymer degrades in cells with the rate that does not cells or tissues. Thus a biocompatible group, as used herein, exceed the rate of metabolization or excretion of polymer refers to an aliphatic, cycloalkyl, heteroaliphatic, heterocy fragments by the cells. In preferred embodiments, the poly cloalkyl, aryl, or heteroaryl moiety, which falls within the mers and polymer biodegradation byproducts are biocompat definition of the term biocompatible, as defined above and ible. herein. The term “Biocompatibility” as used herein, is also (0203 “Bioavailability': The term “bioavailability” refers taken to mean that the compounds exhibit minimal interac to the systemic availability (i.e., blood/plasma levels) of a tions with recognition proteins, e.g., naturally occurring anti given amount of drug or compound administered to a Subject. bodies, cell proteins, cells and other components of biological Bioavailability is an absolute term that indicates measure systems, unless Such interactions are specifically desirable. ment of both the time (rate) and total amount (extent) of drug Thus, Substances and functional groups specifically intended or compound that reaches the general circulation from an to cause the above minimal interactions, e.g., drugs and pro administered dosage form. drugs, are considered to be biocompatible. Preferably (with (0204 “Hydrophilic'. The term “hydrophilic” as it relates exception of compounds intended to be cytotoxic, Such as, to Substituents on the polymer monomeric units does not e.g., antineoplastic agents), compounds are “biocompatible” essentially differ from the common meaning of this term in if their addition to normal cells in vitro, at concentrations the art, and denotes chemical moieties which contain ioniz similar to the intended systemic in Vivo concentrations, able, polar, or polarizable atoms, or which otherwise may be results in less than or equal to 1% cell death during the time Solvated by water molecules. Thus a hydrophilic group, as equivalent to the half-life of the compound in vivo (e.g., the used herein, refers to an aliphatic, cycloalkyl, heteroaliphatic, period of time required for 50% of the compound adminis heterocycloalkyl, aryl or heteroaryl moiety, which falls tered in vivo to be eliminated/cleared), and their administra within the definition of the term hydrophilic, as defined tion in vivo induces minimal and medically acceptable above. Examples of particular hydrophilic organic moieties inflammation, foreign body reaction, immunotoxicity, which are suitable include, without limitation, aliphatic or chemical toxicity and/or other such adverse effects. In the heteroaliphatic groups comprising a chain of atoms in a range above sentence, the term “normal cells' refers to cells that are of between about one and twelve atoms, hydroxyl, hydroxy not intended to be destroyed or otherwise significantly alkyl, amine, carboxyl, amide, carboxylic ester, thioester, affected by the compound being tested. aldehyde, nitryl, isonitryl. nitroso, hydroxylamine, mercap (0202) “Biodegradable'. As used herein, “biodegradable” toalkyl, heterocycle, carbamates, carboxylic acids and their polymers are polymers that are Susceptible to biological pro salts, Sulfonic acids and their salts, Sulfonic acid esters, phos cessing in vivo. As used herein, "biodegradable' compounds phoric acids and their salts, phosphate esters, polyglycol or moieties are those that, when taken up by cells, can be ethers, polyamines, polycarboxylates, polyesters and poly broken down by the lysosomal or other chemical machinery thioesters. In preferred embodiments of the present invention, or by hydrolysis into components that the cells can either at least one of the polymer monomeric units include a car reuse or dispose of without significant toxic effect on the boxyl group (COOH), an aldehyde group (CHO), a methylol cells. The term “biocleavable' as used herein has the same (CHOH) or a glycol (for example, CHOH-CH-OH or meaning of "biodegradable'. The degradation fragments CH-(CHOH)). preferably induce little or no organ or cell overload or patho 0205 The term “hydrophilic” as it relates to the polymers logical processes caused by Such overload or other adverse of the invention generally does not differ from usage of this effects in vivo. Examples of biodegradation processes include term in the art, and denotes polymers comprising hydrophilic enzymatic and non-enzymatic hydrolysis, oxidation and functional groups as defined above. In a preferred embodi reduction. Suitable conditions for non-enzymatic hydrolysis ment, hydrophilic polymer is a water-soluble polymer. of the biodegradable protein-polymer-drug conjugates (or Hydrophilicity of the polymer can be directly measured their components, e.g., the biodegradable polymeric carrier through determination of hydration energy, or determined and the linkers between the carrier and the antibody or the through investigation between two liquid phases, or by chro drug molecule) described herein, for example, include expo matography on Solid phases with known hydrophobicity, Sure of the biodegradable conjugates to waterata temperature such as, for example, C4 or C18. and a pH of lysosomal intracellular compartment. Biodegra 0206 “Polymeric Carrier'': The term polymeric carrier, as dation of Some protein-polymer-drug conjugates (or their used herein, refers to a polymer or a modified polymer, which components, e.g., the biodegradable polymeric carrier and is suitable for covalently attaching to or can be covalently the linkers between the carrier and the antibody or the drug attached to one or more drug molecules with a designated molecule), can also be enhanced extracellularly, e.g. in low linker and/or one or more PBRMs with a designated linker. pH regions of the animal body, e.g. an inflamed area, in the 0207 “Physiological conditions: The phrase “physi close vicinity of activated macrophages or other cells releas ological conditions, as used herein, relates to the range of ing degradation facilitating factors. In certain preferred chemical (e.g., pH, ionic strength) and biochemical (e.g., embodiments, the effective size of the polymer carrier at enzyme concentrations) conditions likely to be encountered pH-7.5 does not detectably change over 1 to 7 days, and in the extracellular fluids of living tissues. For most normal remains within 50% of the original polymer size for at least tissues, the physiological pH ranges from about 7.0 to 7.4. several weeks. At pH-5, on the other hand, the polymer Circulating blood plasma and normal interstitial liquid rep carrier preferably detectably degrades over 1 to 5 days, and is resent typical examples of normal physiological conditions. completely transformed into low molecular weight fragments 0208 “Polysaccharide”, “carbohydrate' or "oligosaccha within a two-week to several-month time frame. Polymer ride': The terms “polysaccharide”, “carbohydrate', or “oli integrity in Such tests can be measured, for example, by size gosaccharide' are known in the art and refer, generally, to US 2013/0309192 A1 Nov. 21, 2013 22
Substances having chemical formula (CH2O), where gener invention may be found in “Pharmaceutical Substances: Syn ally n>2, and their derivatives. Carbohydrates are polyhy theses, Patents, Applications” by Axel Kleemann and Jurgen droxyaldehydes or polyhydroxyketones, or change to Such Engel, Thieme Medical Publishing, 1999 and the “Merck Substances on simple chemical transformations, such as Index: An Encyclopedia of Chemicals, Drugs, and Biologi hydrolysis, oxidation or reduction. Typically, carbohydrates cals”, Edited by Susan Budavariet al., CRC Press, 1996, both are present in the form of cyclic acetals or ketals (such as, of which are incorporated herein by reference. In preferred glucose or fructose). These cyclic units (monosaccharides) embodiments, the drug used in this invention is a therapeutic may be connected to each other to form molecules with few agent that has antiproliferative (cytostatic and/or cytotoxic) (oligosaccharides) or several (polysaccharides) monosaccha activity against a target cell or pathway. The drug may have a ride units. Often, carbohydrates with well defined number, chemically reactive group Such as, for example, —COOH, types and positioning of monosaccharide units are called primary amine, secondary amine —NHR, —OH, -SH, oligosaccharides, whereas carbohydrates consisting of mix C(O)H, C(O)R, C(O)NHR'. C(S)OH, -S(O) tures of molecules of variable numbers and/or positioning of OR,-P(O) OR, CN, NC or ONO, in which Ris monosaccharide units are called polysaccharides. The terms an aliphatic, heteroaliphatic, carbocyclic or heterocycloalkyl "polysaccharide', 'carbohydrate', and "oligosaccharide'. moiety and R’ is a hydrogen, an aliphatic, heteroaliphatic, are used herein interchangeably. A polysaccharide may carbocyclic, or heterocyclic moiety. include natural Sugars (e.g., glucose, fructose, galactose, 0212 “Drug derivative' or “modified drug or the like as mannose, arabinose, ribose, and Xylose) and/or derivatives of used herein, refers to a compound that comprises the drug naturally occurring Sugars (e.g. 2'-fluororibose, 2'-deoxyri molecule intended to be delivered by the conjugate of the bose, and hexose). invention and a functional group capable of attaching the drug 0209 “Small molecule: As used herein, the term “small molecule to the polymeric carrier. molecule' refers to molecules, whether naturally-occurring 0213 “Active form as used herein refers to a form of a or artificially created (e.g., via chemical synthesis) that have compound that exhibits intended pharmaceutical efficacy in a relatively low molecular weight. Preferred small molecules Vivo or in vitro. In particular, when a drug molecule intended are biologically active in that they produce a local or systemic to be delivered by the conjugate of the invention is released effect in animals, preferably mammals, more preferably from the conjugate, the active form can be the drug itselfor its humans. In certain preferred embodiments, the Small mol derivatives, which exhibit the intended therapeutic proper ecule is a drug and the Small molecule is referred to as "drug ties. The release of the drug from the conjugate can be molecule' or "drug or “therapeutic agent”. In certain achieved by cleavage of a biodegradable bond of the linker embodiments, the drug molecule has MW less than or equal which attaches the drug to the polymeric carrier. The active to about 5 kDa. In other embodiments, the drug molecule has drug derivatives accordingly can comprise a portion of the MW less than or equal to about 1.5kDa. In embodiments, the linker. drug molecule is selected from Vinca alkaloids, auristatins, 0214) “Diagnostic label: As used herein, the term diag tubulysins, duocarmycins, kinase inhibitors, MEK inhibitors, nostic label refers to an atom, group of atoms, moiety or KSP inhibitors, and analogs thereof. Preferably, though not functional group, a nanocrystal, or other discrete element of a necessarily, the drug is one that has already been deemed safe composition of matter, that can be detected in vivo or ex vivo and effective for use by an appropriate governmental agency using analytical methods known in the art. When associated or body, e.g., the FDA. For example, drugs for human use with a conjugate of the present invention, such diagnostic listed by the FDA under 21 C.F.R. SS330.5,331 through 361, labels permit the monitoring of the conjugate in vivo. Alter and 440 through 460; drugs for veterinary use listed by the natively or additionally, constructs and compositions that FDA under 21 C.F.R. SS500 through 589, incorporated herein include diagnostic labels can be used to monitor biological by reference, are all considered suitable for use with the functions or structures. Examples of diagnostic labels present hydrophilic polymers. include, without limitation, labels that can be used in medical 0210 Classes of drug molecules that can be used in the diagnostic procedures, such as, radioactive isotopes (radio practice of the present invention include, but are not limited nuclides) for gamma Scintigraphy and Positron Emission to, anti-cancer Substances, radionuclides, vitamins, anti Tomography (PET), contrast agents for Magnetic Resonance AIDS Substances, antibiotics, immunosuppressants, anti-vi Imaging (MRI) (for example paramagnetic atoms and Super ral Substances, enzyme inhibitors, neurotoxins, opioids, hyp paramagnetic nanocrystals), contrast agents for computed notics, anti-histamines, lubricants, tranquilizers, anti tomography and other X-ray-based imaging methods, agents convulsants, muscle relaxants and anti-Parkinson Substances, for ultrasound-based diagnostic methods (Sonography), anti-spasmodics and muscle contractants including channel agents for neutron activation (e.g., boron, gadolinium), fluo blockers, miotics and anti-cholinergics, anti-glaucoma com rophores for various optical procedures, and, in general moi pounds, anti-parasite and/or anti-protozoal compounds, eties which can emit, reflect, absorb, scatter or otherwise modulators of cell-extracellular matrix interactions including affect electromagnetic fields or waves (e.g. gamma-rays, cell growth inhibitors and anti-adhesion molecules, vasodi X-rays, radiowaves, microwaves, light), particles (e.g. alpha lating agents, inhibitors of DNA, RNA or protein synthesis, particles, electrons, positrons, neutrons, protons) or other anti-hypertensives, analgesics, anti-pyretics, Steroidal and forms of radiation, e.g. ultrasound. non-steroidal anti-inflammatory agents, anti-angiogenic fac 0215 “Aliphatic'. In general, the term aliphatic, as used tors, anti-secretory factors, anticoagulants and/or antithrom herein, includes both saturated and unsaturated, straight chain botic agents, local anesthetics, ophthalmics, prostaglandins, (i.e., unbranched) or branched aliphatic hydrocarbons, which anti-depressants, anti-psychotic Substances, anti-emetics, are optionally Substituted with one or more functional groups. imaging agents. Many large molecules are also drugs. As will be appreciated by one of ordinary skill in the art, 0211. A more complete, although not exhaustive, listing of “aliphatic' is intended herein to include, but is not limited to, classes and specific drugs suitable for use in the present alkyl, alkenyl, alkynyl moieties. Thus, as used herein, the US 2013/0309192 A1 Nov. 21, 2013
term “alkyl includes straight and branched alkyl groups. An phatic chain which contains one or more oxygen, Sulfur, analogous convention applies to other generic terms such as nitrogen, phosphorus or silicon atoms, e.g., in place of carbon “alkenyl', 'alkynyl and the like. Furthermore, as used atoms. Heteroaliphatic moieties may be branched or linear herein, the terms “alkyl”, “alkenyl', 'alkynyl and the like unbranched. In certain embodiments, heteroaliphatic moi encompass both Substituted and unsubstituted groups. In cer eties are substituted (“substituted heteroaliphatic') by inde tain embodiments, as used herein, “lower alkyl is used to pendent replacement of one or more of the hydrogen atoms indicate those alkyl groups (Substituted, unsubstituted, thereon with one or more moieties including, but not limited branched or unbranched) having about 1-6 carbon atoms. to aliphatic; heteroaliphatic; cycloalkyl; heterocycloalkyl; 0216 “Alkenyl: the term alkenyl denotes a monovalent aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy: aryloxy; group derived from a hydrocarbon moiety having at least one heteroalkoxy; heteroaryloxy; alkylthio: arylthio; heteroalky carbon-carbon double bond by the removal of a single hydro lthio; heteroarylthio; F. Cl; Br; I; N: —CN; —C: gen atom. “Substituted alkenyl groups are substituted with one or more functional groups. Substituents include, but are not limited to, any of the substituents mentioned below, i.e., the substituents recited below resulting in the formation of a stable compound. Alkenyl groups include, for example, ethe nyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like. 0217 “Alkynyl: the term alkynyl as used herein refers to a monovalent group derived from a hydrocarbon having at least one carbon-carbon triple bond by the removal of a single hydrogen atom. “Substituted alkenyl groups are substituted —SN' , wherein each occurrence of RG1, RG2 and with one or more functional groups. Substituents include, but RG3 independently includes, but is not limited to, hydrogen, are not limited to, any of the substituents mentioned below, halogen, or an optionally Substituted aliphatic, het i.e., the substituents recited below resulting in the formation eroaliphatic, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, of a stable compound. Representative alkynyl groups include alkylaryl, or alkylheteroaryl moiety. Additional examples of ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like. generally applicable Substituents are illustrated by the spe 0218. In certain embodiments, the alkyl, alkenyl and alky cific embodiments shown in the Examples that are described nyl groups employed in the invention contain about 1-20 herein. aliphatic carbon atoms. In certain other embodiments, the 0221) “Cycloalkyl”: as used herein, the term cycloalkyl alkyl, alkenyl, and alkynyl groups employed in the invention refers to a saturated or unsaturated nonaromatic hydrocarbon contain about 1-10 aliphatic carbon atoms. In yet other mono- or multi-ring system having 3 to 30 carbonatoms (e.g., embodiments, the alkyl, alkenyl, and alkynyl groups C-C). Suitable cycloalkyls include, but are not limited to employed in the invention contain about 1-8 aliphatic carbon cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohep atoms. In still other embodiments, the alkyl, alkenyl, and tyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, alkynyl groups employed in the invention contain about 1-6 cycloheptynyl, adamantyl, and the like. aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention con 0222 “Heterocycloalkyl as used herein refers to a satu tain about 1-4 carbonatoms. Illustrative aliphatic groups thus rated or unsaturated nonaromatic 3-8 membered monocyclic, include, but are not limited to, for example, methyl, ethyl, 8-12 membered bicyclic, or 11-19 membered tricyclic ring n-propyl, isopropyl, allyl. n-butyl, sec-butyl, isobutyl, tert system having one or more heteroatoms (such as O, N, S, or butyl, n-pentyl, sec-pentyl, isopentyl, tert-pentyl, n-hexyl, Se), unless specified otherwise. In certain embodiments, the sec-hexyl, moieties and the like, which again, may bear one or term "heterocycloalkyl refers to a non-aromatic 5-, 6-, 7- or more Substituents. Alkenyl groups include, but are not limited 8-membered ring or a polycyclic group, including, but not to, for example, ethenyl, propenyl, butenyl, 1-methyl-2- limited to a bi- or tri-cyclic group comprising fused six buten-1-yl, and the like. Representative alkynyl groups membered rings having between one and three heteroatoms include, but are not limited to, ethynyl, 2-propynyl (propar independently selected from oxygen, Sulfur and nitrogen, gyl), 1-propynyl and the like. wherein (i) each 5-membered ring has 0 to 2 double bonds and 0219 “Alkylene' as used herein, the term alkylene by each 6-membered ring has 0 to 2 double bonds, (ii) the nitro itself or part of another term refers to a saturated, branched or gen and Sulfur heteroatoms may optionally be oxidized, (iii) straight chain having two monovalent radical centers derived the nitrogen heteroatom may optionally be quaternized, and by the removal of two hydrogenatoms from the same or two (iv) any of the above heterocycloalkyl; rings may be fused to different carbon atoms of a parent alkane. Alkylene radicals an aryl or heteroaryl ring. Examples of heterocycloalkyl include, but are not limited to, methylene, 1.2, ethylene, 1,3- groups include, but are not limited to, piperidinyl, piperazi propyl, and the like. Suitable alkylenes include, but are not nyl, pyrrolidinyl, dioxanyl, tetrahydrofuranyl, tetrahy limited to methylene, ethylene, propylene, butylene, penty drothienyl, isolindolinyl, indolinyl, imidazolidinyl, pyrazo lene, hexylene, heptylene, ocytylene, nonylene, decalene, lidinyl, oxazolidinyl, isoxazolidinyl, triazolidinyl, and the like. The term “cycloalkylene similarly refers to tetrahyrofuranyl, oxiranyl, aZetidinyl, oxetanyl, thietanyl. bivalent cycloalkyl. Cycloalkylene radicals include, but are 1.2.3,6-tetrahydropyridinyl, tetrahydro-2H-pyranyl. 3,6-di not limited to, 11-cyclopentylene, 1.2-cyclopentylene, 1,1- hydro-2H-pyranyl, morpholinyl, and the like. cyclobutylene, 1.3-cyclobutylene, etc. 0223) Aryl’: as used herein, refers to groups with aroma 0220 “Heteroaliphatic': as used herein, the term het ticity, including "conjugated or multicyclic systems with at eroaliphatic refers to aliphatic moieties in which one or more least one aromatic ring and do not contain any heteroatom in carbon atoms in the main chain have been substituted with a the ring structure. Examples include phenyl, benzyl, 1.2.3,4- heteroatom. Thus, a heteroaliphatic group refers to an ali tetrahydronaphthalenyl, etc. US 2013/0309192 A1 Nov. 21, 2013 24
0224) “Heteroaryl': as used herein, refers to aryl groups, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, as defined above, except having from one to four heteroatoms benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carba in the ring structure, and may also be referred to as “aryl Zolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, heterocycles' or "heteroaromatics.” As used herein, the term cinnolinyl, decahydroquinolinyl, 2H,6H-1.5.2-dithiazinyl, "heteroaryl is intended to include a stable 5-, 6-, or 7-mem dihydrofuro2,3-btetrahydrofuran, furanyl, furazanyl, imi bered monocyclic or 7- 8-, 9-, 10-, 11- or 12-membered dazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indole bicyclic aromatic heterocyclic ring which consists of carbon nyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, atoms and one or more heteroatoms, e.g., 1 or 1-2 or 1-3 or 1-4 isobenzofuranyl, isochromanyl, isolindazolyl, isoindolinyl, or 1-5 or 1-6 heteroatoms, or e.g., 1, 2, 3, 4, 5, or 6 heteroa isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methyl toms, independently selected from the group consisting of enedioxyphenyl, morpholinyl, naphthyridinyl, octahydroiso nitrogen, oxygen and Sulfur. The nitrogen atom may be Sub quinolinyl, oxadiazolyl, 1.2.3-oxadiazolyl, 1,2,4-oxadiaz stituted or unsubstituted (i.e., Nor NR wherein R is Horother olyl, 1,2,5-oxadiazolyl, 1.3,4-oxadiazolyl, 1,2,4-oxadiazols Substituents, as defined). The nitrogen and Sulfur heteroatoms (4H)-one, oxazolidinyl, oxazolyl, OXindolyl, pyrimidinyl, may optionally be oxidized (i.e., N->O and S(O), where p=1 phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazi or 2). It is to be noted that total number of S and Oatoms in the nyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, aromatic heterocycle is not more than 1. Examples of het piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, eroaryl include pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyra pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, tetrazolyl, Zolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridot oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, hiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrroli furanyl, quinolinyl, isoquinolinyl, tetrazolyl pyridazinyl, nyl, 2H-pyrrolyl pyrrolyl, quinazolinyl, quinolinyl, quinazolinyl, dihydroquinazolyl, and tetrahydroquinazolyl 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofura and the like. nyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetra 0225. Furthermore, the terms “aryl” and “heteroaryl Zolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thia include multicyclic aryland heteroaryl groups, e.g., tricyclic, diazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, bicyclic, e.g., naphthalene, benzoxazole, benzodioxazole, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimi benzothiazole, benzoimidazole, benzothiophene, methylene dazolyl, thiophenyl, triazinyl, 1.2.3-triazolyl, 1,2,4-triazolyl, dioxyphenyl, quinoline, isoquinoline, naphthrydine, indole, 1,2,5-triazolyl, 1,3,4-triazolyl and Xanthenyl. Multiple-ring benzofuran, purine, benzofuran, deaZapurine, indolizine. heterocycle can include fused, bridged or spiro rings. 0226. In the case of multicyclic aromatic rings, only one of 0230. The cycloalkyl, heterocycloalkyl, aryl, or heteroaryl the rings needs to be aromatic (e.g., 2,3-dihydroindole), ring (or the carbocyclic or heterocyclic group) can be substi although all of the rings may be aromatic (e.g., quinoline). tuted at one or more ring positions (e.g., the ring-forming The second ring can also be fused or bridged. carbon or heteroatom such as N) with such substituents as 0227 “Carbocycle” or “carbocyclic moiety” as used described above, for example, aliphatic; heteroaliphatic; herein, is intended to include any stable monocyclic, bicyclic cycloalkyl; heterocycloalkyl, aryl; heteroaryl; alkylaryl; or tricyclic ring having the specified number of carbons, any alkylheteroaryl; alkoxy: aryloxy; heteroalkoxy; heteroary of which may be saturated, unsaturated, or aromatic. Car loxy; alkylthio: arylthio; heteroalkylthio; heteroarylthio; F: bocycle includes cycloalkyl and aryl. For example, a C-C, Cl; Br; I; NO; CN; CF; CHCF; —CHCl: carbocycle is intended to include a monocyclic, bicyclic or —CHOH, -CHCH-OH, -CH-NH; —CHSOCH : tricyclic ring having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms. Examples of carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopen tyl, cyclopentenyl, cyclohexyl, cycloheptenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, cyclooctadienyl, fluorenyl, phenyl, naphthyl, indanyl, adamanty1 and tetrahy dronaphthyl. Bridged rings are also included in the definition of carbocycle, including, for example, 3.3.0 bicyclooctane, 4.3.0 bicyclononane, 4.4.0 bicyclodecane and 2.2.2 bicy —SONR' , wherein each occurrence of R', R'' and clooctane. A bridged ring occurs when one or more carbon R’ independently includes, but is not limited to, hydrogen, atoms link two non-adjacent carbon atoms. In one embodi halogen, or an optionally Substituted aliphatic, het ment, bridge rings are one or two carbonatoms. It is noted that eroaliphatic, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, a bridge always converts a monocyclic ring into a tricyclic alkylaryl, or alkylheteroaryl moiety. Aryl and heteroaryl ring. When a ring is bridged, the substituents recited for the groups can also be fused or bridged with cycloalkyl or het ring may also be present on the bridge. Fused (e.g., naphthyl, erocyclic rings, which are not aromatic so as to form a mul tetrahydronaphthyl) and spiro rings are also included. ticyclic system (e.g., tetralin, methylenedioxyphenyl). 0228 “Heterocycle” or "heterocyclic moiety” as used 0231 “Alkoxy' (or “alkyloxy): as used herein, the term herein, includes any ring structure (saturated, unsaturated, or alkoxy (or alkyloxy) refers to an alkyl group, as previously aromatic) which contains at least one ring heteroatom (e.g., defined, attached to the parent molecular moiety through an N, O or S). Heterocycle includes heterocycloalkyl and het oxygen atom ("alkoxy”). In certain embodiments, the alkyl eroaryl. Examples of heterocycles include, but are not limited group contains about 1-20 aliphatic carbon atoms. In certain to, morpholine, pyrrolidine, tetrahydrothiophene, piperidine, other embodiments, the alkyl group contains about 1-10 ali piperazine and tetrahydrofuran. phatic carbon atoms. In yet other embodiments, the alkyl 0229. Examples of heterocyclic groups include, but are group contains about 1-8 aliphatic carbonatoms. In still other not limited to, acridinyl, azocinyl, benzimidazolyl, benzo embodiments, the alkyl group contains about 1-6 aliphatic furanyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, carbon atoms. In yet other embodiments, the alkyl group US 2013/0309192 A1 Nov. 21, 2013 contains about 1-4 aliphatic carbon atoms. Examples of a Sulfur atom which is covalently bonded to an alkenyl group; alkoxy groups, include but are not limited to, methoxy, and alkthioalkynyls' refers to moieties wherein an alkyl, ethoxy, propoxy, isopropoxy, n-butoxy, tert-butoxy, neopen alkenyl or alkynyl group is bonded to a Sulfur atom which is toxy and n-hexoxy. covalently bonded to an alkynyl group. 0232 'Aryloxy’: as used herein, the termaryloxy refers to 0239 Arylthio” (or “thioaryl’) means an aryl group as an aryl group, as defined herein, attached to the parent defined herein with the indicated number of carbon atoms molecular moiety through an oxygenatom. Examples of ary attached through a Sulfur atom. loxy groups include but are not limited to phenoxy and 0240 “Carboxylic acid' as used herein refers to a com napthyloxy. pound comprising a group of formula -CO2H. 0233 “Heteroaryloxy’: as used herein, the term het 0241 “Dicarboxylic acid refers to a compound compris eroaryloxy refers to a heteroaryl group, as defined herein, ing two groups of formula —COH. attached to the parent molecular moiety through an oxygen 0242 "Halo, halide and halogen': The terms halo, halide atom. Examples of heteroaryloxy groups include but are not and halogen as used herein refer to an atom selected from limited to, quinolyloxy and isoquinolizinyloxy. fluorine, chlorine, bromine, and iodine. 0234 Amine': the term amine refers to a group having 0243 “Methylol’: The term methylolas used herein refers the structure —N(R) wherein each occurrence of R is inde to an alcohol group of the structure —CH2OH. pendently hydrogen, oran aliphatic or heteroaliphatic moiety, 0244 “Hydroxyalkyl': As used herein, the term hydroxy or the R groups, taken together, may form a heterocyclic alkyl refers to an alkyl group, as defined above, bearing at moiety. In certain instances, an amine group can be charged least one OH group. (protonized) or quarternized, e.g., —HN (R) or - N'(R) 0245 "Mercaptoalkyl': The term mercaptoalkyl as used 0235 Alkylamino': as used herein, the term alkylamino therein refers to an alkyl group, as defined above, bearing at refers to a group having the structure —NHR' wherein R' is least one SH group alkyl, as defined herein. The term “aminoalkyl refers to a 0246 Acyl' includes moieties that contain the acyl radi group having the structure NHR' , wherein R' is alkyl, as cal ( C(O) ) or a carbonyl group. “Substituted acyl defined herein. In certain embodiments, the alkyl group con includes acyl groups where one or more of the hydrogen tains about 1-20 aliphatic carbon atoms. In certain other atoms are replaced by, for example, alkyl groups, alkynyl embodiments, the alkyl group contains about 1-10 aliphatic groups, halogen, hydroxyl, alkylcarbonyloxy, arylcarbony carbon atoms. In yet other embodiments, the alkyl, alkenyl, loxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate, and alkynyl groups employed in the invention contain about alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbo 1-8 aliphatic carbon atoms. In still other embodiments, the nyl, alkylaminocarbonyl, dialkylaminocarbonyl, alkylthio alkyl group contains about 1-6 aliphatic carbon atoms. In yet carbonyl, alkoxyl, phosphate, phosphonato, phosphinato, other embodiments, the alkyl group contains about 1-4 ali amino (including alkylamino, dialkylamino, arylamino, dia phatic carbonatoms. Examples of alkylamino include, but are rylamino and alkylarylamino), acylamino (including alkyl not limited to, methylamino, ethylamino, iso-propylamino carbonylamino, arylcarbonylamino, carbamoyl and ureido), and the like. amidino, imino, Sulfhydryl, alkylthio, arylthio, thiocarboxy 0236 “Alkylthio” (or “thioalkyl) means an alkyl group as late, Sulfates, alkylsulfinyl, Sulfonato, Sulfamoyl, Sulfona defined herein with the indicated number of carbon atoms mido, nitro, trifluoromethyl, cyano, azido, heterocyclyl, alky attached through a sulfur atom. C. alkylthio, is intended to laryl, or an aryl or heteroaryl moiety. include C, C2, Cs, Ca, Cs, and C alkylthio groups. Cls 0247 “Hydrocarbon': The term hydrocarbon, as used alkylthio, is intended to include C, C, C, C, Cs, C. C.7. herein, refers to any chemical group comprising hydrogen and Cs alkylthio groups. The thioalkyl groups can be substi and carbon. The hydrocarbon may be substituted or unsub tuted with groups such as alkyl, alkenyl, alkynyl, halogen, stituted. The hydrocarbon may be unsaturated, Saturated, hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbo branched, unbranched, cyclic, polycyclic, or heterocyclic. nyloxy, aryloxycarbonyloxy, carboxylate, carboxyacid, alky Illustrative hydrocarbons include, for example, methyl, ethyl, lcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl, n-propyl, iso-propyl, cyclopropyl, allyl, vinyl. n-butyl, tert alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbo butyl, ethynyl, cyclohexyl, methoxy, diethylamino, heterocy nyl, alkoxyl, amino (including alkylamino, dialkylamino, cloalkyl, aryl, heteroaryl, thioalkyl, and the like. As would be arylamino, diarylamino and alkylarylamino), acylamino (in known to one skilled in this art, all valencies must be satisfied cluding alkylcarbonylamino, arylcarbonylamino, carbamoyl in making any Substitutions. and ureido), amidino, imino, Sulfhydryl, alkylthio, arylthio. 0248 “Alkylaryl as used herein refers to an aryl group thiocarboxylate, Sulfates, alkylsulfinyl, Sulfonato, Sulfamoyl, Substituted with one or more alkyl groups (e.g., methylphe Sulfonamido, nitro, trifluoromethyl, cyano, azido, heterocy nyl). clyl alkylaryl, or an aryl or heteroaryl moieties. 0249 “Alkylarylamino” as used herein refers to 0237 "Thiocarbonyl' or “thiocarboxy” includes com - NR'R'', wherein R is alkyl, as defined herein, and R' pounds and moieties which contain a carbon connected with is an aryl, as defined herein, or at least one of RandR is a double bond to a sulfur atom. an alkylaryl as defined herein. 0238 “Thioether includes moieties which contain a sul (0250 “Substituted: The terms substituted, whether pre fur atom bonded to two carbon atoms or heteroatoms. ceded by the term “optionally’ or not, and substituent, as used Examples of thioethers include, but are not limited to alkthio herein, refers to the replacement of hydrogen radicals in a alkyls, alkthioalkenyls and alkthioalkynyls. The term “alk given structure with the radical of a specified substituent. thioalkyls' include moieties with an alkyl, alkenyl or alkynyl When more than one position in any given structure may be group bonded to a sulfur atom which is bonded to an alkyl substituted with more than one substituent selected from a group. Similarly, the term “alkthioalkenyls' refers to moi specified group, the Substituent may be either the same or eties wherein an alkyl, alkenyl or alkynyl group is bonded to different at every position. As used herein, the term “substi US 2013/0309192 A1 Nov. 21, 2013 26 tuted” is contemplated to include all permissible substituents L-amino acyl residues. More generally, the unnatural amino of organic compounds. In a broad aspect, the permissible acid comprises a residue of the general formula Substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non aromatic Substituents of organic compounds. Heteroatoms Such as nitrogen may have hydrogen Substituents and/or any permissible Substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. Examples of substituents include, but are not limited to ali /A phatic; heteroaliphatic; cycloalkyl; heterocycloalkyl, aryl; heteroaryl; alkylaryl; alkylheteroaryl; alkoxy: aryloxy; het wherein the side chain R is other than the amino acid side eroalkoxy; heteroaryloxy; alkylthio: arylthio; heteroalky chains occurring in nature. Exemplary unnatural amino acids, lthio; heteroarylthio; F: Cl; Br; I; NO; CN; –CF: include, but are not limited to, sarcosine (N-methylglycine), citrulline (cit), homocitrulline, B-ureidoalanine, thiocitrul line, hydroxyproline, allothreonine, pipecolic acid (ho moproline), C-aminoisobutyric acid, tert-butylglycine, tert butylalanine, allo-isoleucine, norleucine, C.-methyleucine, cyclohexylglycine, B-cyclohexylalanine, B-cyclopentylala nine, C.-methylproline, phenylglycine, C.-methylphenylala nine and homophenylalanine. 0256 Amino acyl: More generally, the term amino acyl, as used herein, encompasses natural amino acid and unnatural NR'SONR , or -SONR' , wherein each amino acids. occurrence of R', RandR independently includes, but (0257 “Polyamide': refers to homo- or hetero-polymers of is not limited to, hydrogen, halogen, or an optionally Substi natural amino acid and unnatural amino acids. Illustrative tuted aliphatic, heteroaliphatic, cycloalkyl, heterocycloalkyl, homo-polymers include, but are not limited to, poly-lysine, aryl, heteroaryl, alkylaryl, or alkylheteroaryl moiety. Addi poly-arginine, poly-Y-glutaric acid, and the like. Illustrative tional examples of generally applicable Substituents are illus hetero-polymers include, but are not limited to, polymers trated by the specific embodiments shown in the Examples comprising peptides fragments selected from peptidases, that are described herein. lysozymes, metalloproteinases, and the like. 0251. The following are more general terms used through (0258 “PHF refers to poly(1-hydroxymethylethylene out the present application: hydroxymethyl-formal). 0252 “Animal’: The term animal, as used herein, refers to humans as well as non-human animals, at any stage of devel 0259. As used herein, the terms “polymer unit”, “mono opment, including, for example, mammals, birds, reptiles, meric unit, "monomer, "monomer unit”, “unit all refer to amphibians, fish, worms and single cells. Cell cultures and a repeatable structural unit in a polymer. live tissue samples are considered to be pluralities of animals. 0260 The present invention is intended to include all iso Preferably, the non-human animal is a mammal (e.g., a topes of atoms occurring in the present compounds. Isotopes rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a include those atoms having the same atomic number but primate, or a pig). An animal may be a transgenic animal or a different mass numbers. By way of general example and human clone. The term 'subject’ encompasses animals. without limitation, isotopes of hydrogen include tritium and 0253) “Efficient amount’: In general, as it refers to an deuterium. Isotopes of carbon include C-13 and C-14. active agent or drug delivery device, the term “efficient 0261 The present invention is intended to include all iso amount” refers to the amount necessary to elicit the desired mers of the compound, which refers to and includes, optical biological response. As will be appreciated by those of ordi isomers, and tautomeric isomers, where optical isomers nary skill in this art, the efficient amount of an agent or device include enantiomers and diastereomers, chiral isomers and may vary depending on Such factors as the desired biological non-chiral isomers, and the optical isomers include isolated endpoint, the agent to be delivered, the composition of the optical isomers as well as mixtures of optical isomers includ encapsulating matrix, the target tissue, etc. For example, the ing racemic and non-racemic mixtures; where an isomer may efficient amount of microparticles containing an antigen to be be in isolated form or in a mixture with one or more other delivered to immunize an individual is the amount that results isomers. in an immune response Sufficient to prevent infection with an organism having the administered antigen. Polymeric Carriers 0254 “Natural amino acid as used herein refers to any 0262. In certain exemplary embodiments, the conjugates one of the common, naturally occurring L-amino acids found of the invention find use in biomedical applications, such as in naturally occurring proteins: glycine (Gly), alanine (Ala), drug delivery and tissue engineering, and the carrier is bio Valine (Val), leucine (Leu), isoleucine (Ile), lysine (LyS), argi compatible and biodegradable. In certain embodiments, the nine (Arg), histidine (His), proline (Pro), serine (Ser), threo carrier is a soluble polymer, nanoparticle, gel, liposome, nine (Thr), phenylalanine (Phe), tyrosine (Tyr), tryptophan micelle, Suture, implant, etc. In certain embodiments, the (Trp), aspartic acid (Asp), glutamic acid (Glu), asparagine term “soluble polymer encompasses biodegradable biocom (ASn), glutamine (Gln), cysteine (CyS) and methionine (Met). patible polymer Such as a polyal (e.g., hydrophilic polyacetal 0255 “Unnatural amino acid as used herein refers to any or polyketal). In certain other embodiments, the carrier is a amino acid which is not a natural amino acid. This includes, fully synthetic, semi-synthetic or naturally-occurring poly for example, amino acids that comprise Cl-, 3-, co-, D-. mer. In certain other embodiments, the carrier is hydrophilic. US 2013/0309192 A1 Nov. 21, 2013 27
0263. In certain exemplary embodiments, the carriers used consisting of agarose, hyluronan, chondroitinsulfate, derma in the present invention are biodegradable biocompatible tanSulfate, keratansulfate, alginic acid and heparin. polyals comprising at least one hydrolysable bond in each 0269. In yet other exemplary embodiments, the polymeric monomer unit positioned within the main chain. This ensures carrier comprises a copolymer of a polyacetal/polyketal and a that the degradation process (via hydrolysis/cleavage of the hydrophilic polymer selected from the group consisting of monomer units) will result in fragmentation of the polymer polyacrylates, polyvinyl polymers, polyesters, polyorthoe conjugate to the monomeric components (i.e., degradation), sters, polyamides, polypeptides, and derivatives thereof. and confers to the polymer conjugates of the invention their 0270. In yet another embodiment, the polymeric carrier is biodegradable properties. The properties (e.g., Solubility, bio dextrin that is produced by the hydrolysis of a starch obtained adhesivity and hydrophilicity) of biodegradable biocompat from various natural products Such as, for example, wheat, ible polymer conjugates can be modified by Subsequent Sub rice, maize and tapioca. Depending on the structure of the stitution of additional hydrophilic or hydrophobic groups. starch starting material each dextrin comprises a unique dis Examples of biodegradable biocompatible polymers suitable tribution of C-1.4 linkages and C-1,6 linkages. Since the rate for practicing the invention can be found interalia in U.S. Pat. of biodegradability of C-1,6 linkages is typically less than that Nos. 5,811,510; 5,863,990; 5,958,398; 7,838,619 and 7,790, for C-14 linkages, preferably the percentage of C-1,6 link 150; and U.S. Publication No. 2006/0058512; each of the ages is less than 10% and more preferably less than 5%. In one above listed patent documents is incorporated herein by ref embodiment the molecular weight of the dextrin is in the erence in its entirety. Guidance on the significance, prepara range of about 1 kDa to about 200 kDa, more preferably from tion, and applications of this type of polymers may be found about 2 kDa to about 55 kDa. in the above-cited documents. In certain embodiments, it is 0271 In certain embodiments, the carrier comprises anticipated that the present invention will be particularly use polysaccharides activated by selective oxidation of cyclic ful in combination with the above-referenced patent docu vicinal diols of 1.2-, 1.4-, 1.6-, and 2,6-pyranosides, and 1.2-, ments, as well as U.S. Pat. Nos. 5,582,172 and 6,822,086, 1.5-, 1.6-furanosides, or by oxidation of lateral 6-hydroxy each of the above listed patent documents is incorporated and 5,6-diol containing polysaccharides prior to conjugation herein by reference in its entirety. with drug molecules or PBRMs. 0264. The conjugates of this invention are hydrophilic, 0272. In still other embodiments, the polymeric carrier hydrolysable and comprise drug molecules (e.g., Vinca alka comprises a biodegradable biocompatible polyacetal wherein loids or derivatives, non-natural camptothecin compounds or at least a Subset of the polyacetal repeat structural units have derivatives, auristatins, tubulysins, duocarmycins, PI3 the following chemical structure: kinases, MEK inhibitors, KSP inhibitors, and analogs thereof) and antibodies (e.g., Trastuzumab, Cetuximab, Rit uximab, Bevacizumab, Epratuzumab, Veltuzumab, Labetu R R3 Rs Zumab) or peptides (LHRH receptor targeting peptides, EC-1 peptide) covalently attached to the polymer carrier via link --O-C-O-C-R-H ages that contain one or more biodegradable bonds. Thus, in certain exemplary embodiments, carriers suitable for practic ing the present invention are polyals having at least one acetal/ketal oxygen atom in each monomer unit positioned 0273 wherein for each occurrence of the n bracketed within the main chain. As discussed above, this ensures that structure, one of R and R is hydrogen, and the other is a the degradation process (via hydrolysis/cleavage of the poly biocompatible group and includes a carbon atom covalently mer acetal/ketal groups) will result in fragmentation of the attached to C'; R is a carbon atom covalently attached to C: polyal conjugate to low molecular weight components (i.e., n" is an integer, each occurrence of R. R. Rs and R is a degradation). biocompatible group and is independently hydrogen or an 0265. In certain embodiments, biodegradable biocompat organic moiety; and for each occurrence of the bracketed ible polymer carriers, used for preparation of polymer conju structure n, at least one of R. R. R. R. Rs and R comprises gates of the invention, are naturally occurring polysaccha a functional group suitable for coupling. In certain embodi rides, glycopolysaccharides, and synthetic polymers of ments, the functional group is a hydroxyl moiety. polyglycoside, polyacetal, polyamide, polyether, and polyes 0274. In one embodiment, the polymeric carrier com ter origin and products of their oxidation, fictionalization, prises activated hydrophilic biodegradable biocompatible modification, cross-linking, and conjugation. polymers comprising from 0.1% to 100% polyacetal moieties 0266. In certain other embodiments, the carrier is a hydro whose backbone is represented by the following chemical philic biodegradable polymer selected from the group con Structure: sisting of carbohydrates, glycopolysaccharides, glycolipids, glycoconjugates, polyacetals, polyketals, and derivatives thereof. Wherein: 0267 In certain exemplary embodiments, the carrier is a naturally occurring linear and/or branched biodegradable (0275 R, and Rs are independently hydrogen, hydroxyl, biocompatible homopolysaccharide selected from the group hydroxy alkyl (e.g., —CH2OH. —CH(OH)—CH(OH), consisting of cellulose, amylose, dextran, levan, fucoidan, - CHO,-CH(OH) CHO or -carbonyl; and carraginan, inulin, pectin, amylopectin, glycogen and lix (0276 o is an integer from 20 to 2000. Cal. 0277. In yet other embodiments, the polymeric carrier 0268. In certain other exemplary embodiments, the carrier comprises a biodegradable biocompatible polyketal wherein is a naturally occurring linear and branched biodegradable at least a subset of the polyketal repeatable structural units biocompatible heteropolysaccharide selected from the group have the following chemical structure: US 2013/0309192 A1 Nov. 21, 2013 28
-continued (B) O O R R3 Rs --O-C-O-C-R -- HO OH R2 R4 R6 or OH (C) wherein each occurrence of R and R is a biocompatible group and R. R. R. Rs. Re and are as defined herein 0278. In certain embodiments, the ketal units are mono HO HO r mers of Formula (IIa) or (IIb): 0281. In another embodiment, the polymeric carrier com prises unmodified acetal units, i.e., polyacetal segments. In (IIa) OH Some embodiments, the polyacetals can be derived from exhaustively oxidized dextran followed by reduction. These O O polymers have been described in U.S. Pat. No. 5,811,510, which is hereby incorporated by reference for its description of polyacetals at column 2, line 65 to column 8, line 55 and OH, OH O their synthesis at column 10, line 45 to column 11, line 14. In (IIb) one embodiment, the unmodified polyacetal polymer is a OH poly(hydroxymethylethylene hydroxymethyl formal) poly mer (PHF). O 0282. In addition to poly(hydroxymethylethylene hydroxymethyl formal) polymers, the backbone of the poly meric carrier can also comprise co-polymers of poly(hy O droxymethylethylene hydroxymethyl formal) blocks and other acetal or non-acetal monomers or polymers. For example, polyethylene glycol polymers are useful as a stealth OH OH agent in the polymer backbone because they can decrease interactions between polymer side chains of the appended functional groups. Such groups can also be useful in limiting 0279 Biodegradable, biocompatible polyketal polymers interactions such as between serum factors and the modified polymer. Other stealth agent monomers for inclusion in the and their methods of making have been described in U.S. Pat. polymer backbone include, for example, ethyleneimine, Nos. 5,811,510, 7,790,150 and 7,838,619, which are hereby methacrylic acid, acrylamide, glutamic acid, and combina incorporated by reference in their entirety. tions thereof. 0280. In one embodiment, the polymeric carrier can be 0283. The acetal or ketal units are present in the modified obtained from partially oxidized dextran (B1->6)-D-glucose) polymer in an amount effective to promote biocompatibility. followed by reduction. In this embodiment, the polymer.com The unmodified acetal or ketal unit can be described as a prises a random mixture of the unmodified dextran (A), par “stealth agent” that provides biocompatibility and solubility tially oxidized dextran acetal units (B) and exhaustively dex to the modified polymers. In addition, conjugation to a poly tran acetal units (C) of the following structures: acetal or polyketal polymer can modify the Susceptibility to metabolism and degradation of the moieties attached to it, and influence biodistribution, clearance and degradation. (A) 0284. The unmodified acetal units are monomers of For O O mula (III):
HO OH (III)
OH (B) c 0285. The molar fraction, n, of unmodified polyacetal HO OH units is the molar fraction available to promote biocompat ibility, solubility and increase half-life, based on the total OH number of polymer units in the modified polymer. The molar fraction n may be the minimal fraction of unmodified mono mer acetal units needed to provide biocompatibility, solubil US 2013/0309192 A1 Nov. 21, 2013 29 ity, stability, or a particular half-life, or can be some larger the glycerol moiety of the units as well as units having two X' fraction. The most desirable degree of cytotoxicity is substan groups (or other substituents such as -LP-D) with one tially none, i.e., the modified polymer is Substantially inert to attached to the glycolaldehyde moiety and the other attached the subject. However, as is understood by those of ordinary to the glycerol moiety of the units. skill in the art, some degree of cytotoxicity can be tolerated 0287. In one embodiment, biodegradable biocompatible depending on the severity of disease or symptom being polyals Suitable for practicing the present invention have a treated, the efficacy of the treatment, the type and degree of molecular weight of between about 0.5 and about 300 kDa. In immune response, and like considerations. a preferred embodiment of the present invention, the biode 0286. In one embodiment, the modified polymer back gradable biocompatible polyals have a molecular weight of bone comprises units of Formula (IV): between about 1 and about 300 kDa (e.g., between about 1 and about 200 kDa, between about 2 and about 300 kDa, between about 2 and about 200 kDa, between about 5 and (IV) about 100kDa, between about 10 and about 70 kDa, between about 20 and about 50 kDa, between about 20 and about 300 kDa, between about 40 and about 150 kDa, between about 50 and about 100 kDa, between about 2 and about 40 kDa, OH X f it's between about 6 and about 20 kDa, or between about 8 and about 15 kDa). wherein X" indicates the substituent for the hydroxyl group of 0288. In one embodiment, the biodegradable biocompat the polymer backbone. As shown in Formula (IV) and the ible polyals Suitable for practicing the present invention are other formulae described herein, each polyacetal unit has a modified before conjugating with a drug or a PBRM. For single hydroxyl group attached to the glycerol moiety of the example, the polyals may contain subunits of linkers LP or unit and an X" group (or another substituent such as -LP-D) L', such as —C(=O) X-(CH), C(=O)— with X attached to the glycolaldehyde moiety of the unit. This is for being CH, O, or NH, and V being an integer from 1 to 6. Table convenience only and it should be construed that the polymer A below provides some examples of the modified polyals having units of Formula (IV) and other formulae described suitable for conjugating with a drug or PBRM or derivatives herein can contain a random distribution of units having a X" thereof. Unless otherwise specified, reference numbers in group (or another substituent such as -LP-D) attached to the Tables A through E below correspond to the Example num glycolaldehyde moiety of the units and those having a single bers described herein; the term “ND” means not determined: X' group (or another substituent such as -LP-D) attached to and X is CH, O, or NH. TABLE A
Polymer Scaffold for for
O HO
Ex1 O 4. 4. OH, OH O OH O ={ NH
O OH US 2013/0309192 A1 Nov. 21, 2013 30
TABLE A-continued
Ref# Polymer Scaffold
X = CH O O O O O O
Ex's" thisOH OH hisOH O thisOH O X=o X=o X X O s HO HN
)=0 )=o X X
O O HO
SH X Ex12= CH, 1. O r O N O r ON O r ON OH OH OH O)=0 OH O)=0 X X
O O HO HN
O US 2013/0309192 A1 Nov. 21, 2013 31
TABLE A-continued
Ref# Polymer Scaffold
X = CH O O O O
it heOH, OH herOH y- hisOH O X O y O
HO HN
HN O
S A S / \
NO
ch'sOH, OH ch OH O ch OH O )=o )=o X X
HO
HN O
H3C S w S / \ US 2013/0309192 A1 Nov. 21, 2013 32
TABLE A-continued
Ref# Polymer Scaffold
erectorX X
HO'- -
OH, OH O OH O OH
X X
O O OH US 2013/0309192 A1 Nov. 21, 2013 33
TABLE A-continued
Ref# Polymer Scaffold
4 OH, OH 4sO OH 2d.O OH O ={ O ={ X X
OH y
NH O
S- S 7 NV
NO
4- OH OH 4 O OH - OH O
OH US 2013/0309192 A1 Nov. 21, 2013 34
TABLE A-continued
Ref# Polymer Scaffold
4'sOH, OH 4thO OH hisOH O O={ )=O X X
OH S.
.S v N S
ON ICa US 2013/0309192 A1 Nov. 21, 2013 35
TABLE A-continued Ref# Polymer Scaffold r r r X
HO
Therapeutic Agents al., Tet Letts., 50:2932-2935 (2009). Pyrrolobenzodiazepines include those described in Denny, Exp. Opin. Ther: Patents., 0289. In certain embodiments, the therapeutic agent is a 10(4):459-474 (2000). Small molecule having a molecular weight preferably sabout 0295 Exemplary microtubule stabilizing and destabiliz 5 kDa, more preferably sabout 4 kDa, more preferably ing agents include taxane compounds, such as paclitaxel, sabout 3 kDa, most preferably sabout 1.5 kDa or sabout 1 docetaxel; maytansinoids, auristatins and analogs thereof, kDa. tubulysin A and B derivatives, Vinca alkaloid derivatives, 0290. In certain embodiments, about 0.1 to about 25% epothilones and cryptophycins. monomers comprise a therapeutic agent, more preferably 0296 Exemplary maytansinoids or maytansinoid analogs about 0.5 to about 20%, more preferably about 1 to about include maytansinol and maytansinol analogs, maytansine or 15%, and even more preferably about 2 to about 10%. DM-1 and DM-4 are those described in U.S. Pat. Nos. 5,208, 0291. The small molecule therapeutic agents used in this 020; 5,416,064; 6,333.410; 6,441,163; 6,716,821; RE39,151 invention (e.g., antiproliferative (cytotoxic and cytostatic) and 7.276,497. In certain embodiments, the cytotoxic agent is agents capable of being linked to a polymer carrier) include a maytansinoid, another group of anti-tubulin agents (Immu cytotoxic compounds (e.g., broad spectrum), angiogenesis noGen, Inc.; see also Chari et al., 1992, Cancer Res. 52:127 inhibitors, cell cycle progression inhibitors, PI3K/m-TOR/ 131), maytansinoids or maytansinoid analogs. Examples of AKT pathway inhibitors, MAPK signaling pathway inhibi Suitable maytansinoids include maytansinol and maytansinol tors, kinase inhibitors, protein chaperones inhibitors, HDAC analogs. Suitable maytansinoids are disclosed in U.S. Pat. inhibitors, PARP inhibitors, Wnt/Hedgehog signaling path Nos. 4424,219; 4,256,746; 4,294,757; 4,307,016; 4,313,946; way inhibitors and RNA polymerase inhibitors. 4,315,929; 4,331,598; 4.361,650; 4,362,663; 4,364,866; 0292 Broad spectrum cytotoxins include, but are not lim 4,450,254; 4,322,348; 4,371,533; 6,333.410; 5,475,092: ited to, DNA-binding or alkylating drugs, microtubule stabi 5,585,499; and 5,846,545. lizing and destabilizing agents, platinum compounds, and 0297 Exemplary auristatins include auristatin E (also topoisomerase I inhibitors. known as a derivative of dolastatin-10), auristatin EB (AEB), 0293 Exemplary DNA-binding or alkylating drugs auristatin EFP (AEFP), monomethyl auristatin E (MMAE), include, CC-1065 and its analogs, anthracyclines (doxorubi monomethyl auristatin F (MMAF), auristatin F and dolasta cin, epirubicin, idarubicin, daunorubicin) and its analogs, tin. Suitable auristatins are also described in U.S. Publication alkylating agents, such as calicheamicins, dactinomycines, Nos. 2003/0083263, 2011/0020343, and 2011/0070248; PCT mitromycines, pyrrolobenzodiazepines, and the like. Application Publication Nos. WO 09/117,531, WO 2005/ 0294 Exemplary CC-1065 analogs include duocarmycin 081711, WO 04/010957; WO 02/088172 and WO01/24763, SA, duocarmycin Cl, duocarmycin C2, duocarmycin B2, and U.S. Pat. Nos. 7,498,298; 6.884,869; 6,323,315; 6,239, DU-86, KW-2189, bizelesin, seco-adozelesin, and those 104; 6,124,431; 6,034,065; 5,780,588; 5,767,237; 5,665,860; described in U.S. Pat. Nos. 5,475,092: 5,595,499; 5,846,545; 5,663,149; 5,635,483; 5,599,902: 5,554,725; 5,530,097; 6,534,660; 6,586,618; 6,756,397 and 7,049,316. Doxorubi 5,521,284: 5,504,191; 5,410,024; 5,138,036; 5,076,973; cin and its analogs include those described in U.S. Pat. No. 4,986,988: 4,978,744; 4.879,278; 4.816,444; and 4,486,414, 6,630,579. Calicheamicins include those described in U.S. the disclosures of which are incorporated herein by reference Pat. Nos. 5,714,586 and 5,739,116. Duocarmycins include in their entirety. those described in U.S. Pat. Nos. 5,070,092; 5,101,038: 0298 Exemplary tubulysin compounds include com 5,187, 186: 6,548,530; 6,660,742; and 7,553,816 B2; and Liet pounds described in U.S. Pat. Nos. 7.816,377: 7.776,814; US 2013/0309192 A1 Nov. 21, 2013 36
7,754,885; U.S. Publication Nos. 2011/0021568; 2010/ 0306 Exemplary PI3K/m-TOR/AKT signaling pathway 004784; 2010/0048490; 2010/00240701:2008/0176958; and inhibitors include phosphoinositide 3-kinase (PI3K) inhibi PCT Application Nos. WO 98/13375; WO 2004/005269; WO tors, GSK-3 inhibitors, ATM inhibitors, DNA-PK inhibitors 2008/138561; WO 2009/002993; WO 2009/055562; WO and PDK-1 inhibitors. 2009/012958; WO 2009/0261.77; WO 2009/134279; WO (0307 Exemplary PI3 kinases are disclosed in U.S. Pat. 2010/033733; WO 2010/034724; WO 2011/017249; WO No. 6,608,053, and include BEZ235, BGT226, BKM120, 2011/057805; the disclosures of which are incorporated by CAL101, CAL263, demethoxyviridin, GDC-0941, GSK615, reference herein in their entirety. IC87114, LY294.002, Palomid 529, perifosine, PF-0469 1502, 0299| Exemplary Vinca alkaloids include Vincristine, Vin PX-866, SAR245408, SAR245409, SF1126, Wortmannin, blastine, vindesine, and navelbine (vinorelbine). Suitable XL 147 and XL765. Vinca alkaloids that can be used in the present invention are 0308 Exemplary AKT inhibitors include, but are not lim also disclosed in U.S. Publication Nos. 2002/0103136 and ited to ATT867. 2010/0305149, and in U.S. Pat. No. 7,303,749 B 1, the dis 0309 Exemplary MAPK signaling pathway inhibitors closures of which are incorporated herein by reference in their include MEK, Ras, JNK, B-Raf and p38 MAPK inhibitors. entirety. 0310 Exemplary MEK inhibitors are disclosed in U.S. 0300 Exemplary epothilone compounds include Pat. No. 7,517,994 and include GDC-0973, GSK1120212, epothilone A, B, C, D, E and F, and derivatives thereof. MSC1936369B, AS703026, RO5126766 and RO4987655, Suitable epothilone compounds and derivatives thereof are PD0325901, AZD6244, AZD 8330 and GDC-0973. described, for example, in U.S. Pat. Nos. 6,956,036; 6,989, 0311 Exemplary B-raf inhibitors include CDC-0879, 450; 6,121,029; 6,117,659; 6,096,757; 6,043,372; 5,969,145; PLX-4032, and SB590885. and 5,886,026; and WO 97/19086: WO 98/08849; WO 0312 Exemplary B p38 MAPK inhibitors include BIRB 98/22461; WO 98/25929; WO 98/38192: WO99/01124; WO 796, LY2228820 and SB 2021.90 99/02514; WO99/03848: WO99/07692; WO99/27890; and 0313 Receptor tyrosine kinases (RTK) are cell surface WO 99/28324; the disclosures of which are incorporated receptors which are often associated with signaling pathways herein by reference in their entirety. stimulating uncontrolled proliferation of cancer cells and 0301 Exemplary cryptophycin compounds are described neoangiogenesis. Many RTKS, which over express or have in U.S. Pat. Nos. 6,680,311 and 6,747,021. mutations leading to constitutive activation of the receptor, 0302) Exemplary platinum compounds include cisplatin have been identified, including, but not limited to, VEGFR, (PLATINOL(R), carboplatin (PARAPLATINR), oxaliplatin EGFR. FGFR, PDGFR, EphR and RET receptor family (ELOXATINE(R), iproplatin, ormaplatin, and tetraplatin. receptors. Exemplary RTK specific targets include ErbB2, 0303 Exemplary topoisomerase I inhibitors include FLT-3, c-Kit, c-Met, HIF. camptothecin, camptothecin, derivatives, camptothecin ana 0314) Exemplary inhibitors of ErbB2 receptor (EGFR logs and non-natural camptothecins, such as, for example, family) include but not limited to AEE788 (NVP-AEE788), CPT-11 (irinotecan), SN-38, topotecan, 9-aminocamptoth BIBW2992, (Afatinib), Lapatinib, Erlotinib (Tarceva), and ecin, rubitecan, gimatecan, karenitecin, silatecan, lurtotecan, Gefitinib (Iressa). exatecan, diflomotecan, belotecan, lurtotecan and 539625. 0315 Exemplary RTK inhibitors targeting more then one Other camptothecin compounds that can be used in the signaling pathway (multitargeted kinase inhibitors) include present invention include those described in, for example, J. AP24534 (Ponatinib) that targets FGFR, FLT-3, VEGFR Med. Chem. 29:2358-2363 (1986); J. Med. Chem., 23:554 PDGFR and Bcr-Abl receptors; ABT-869 (Linifanib) that (1980); J. Med. Chem., 30:1774 (1987). targets FLT-3 and VEGFR-PDGFR receptors: AZD2171 that 0304 Angiogenesis inhibitors include, but are not limited, targets VEGFR-PDGFR, Flt-1 and VEGF receptors: CHR MetAP2 inhibitors. Exemplary MetAP2 inhibitors include 258 (Dovitinib) that targets VEGFR-PDGFR, FGFR, Flt-3, fumagillol analogs, meaning any compound that includes the and c-Kit receptors. fumagillin core structure, including fumagillamine, that 0316 Exemplary protein chaperon inhibitors include inhibits the ability of MetAP-2 to remove NH-terminal HSP90 inhibitors. Exemplary HSP90 inhibitors include methionines from proteins as described in Rodeschini et al., J. 17AAG derivatives, BIIB021 BIIB028, SNX-5422, NVP Org. Chem., 69,357-373, 2004 and Liu, et al., Science 282, AUY-922 and KW-2478. 1324-1327, 1998. Nonlimiting examples of “fumagillol ana 0317 Exemplary HDAC inhibitors include Belinostat logs are disclosed in J. Org. Chem., 69,357, 2004, J. Org. (PXD101), CUDC-101, Droxinostat, ITF2357 (Givinostat, Chem., 70, 6870, 2005; European Patent Application 0354 Gavinostat),JNJ-26481585, LAQ824 (NVP-LAQ824, Daci 787, J. Med. Chem., 49, 5645, 2006, Bioorg. Med. Chem., 11, nostat), LBH-589 (Panobinostat), MC1568, MGCD0103 5051, 2003, Bioorg. Med. Chem., 14,91, 2004, Tet. Lett. 40, (Mocetinostat), MS-275 (Entinostat), PCI-24781, Pyroxam 4797, 1999; WO99/61432; U.S. Pat. Nos. 6,603,812; 5,789, ide (NSC 696085), SB939, Trichostatin A and Vorinostat 405; 5,767,293; 6,566,541; and 6,207,704. (SAHA). 0305 Exemplary cell cycle progression inhibitors include 0318 Exemplary PARP inhibitors include iniparib (BSI CDK inhibitors such as, for example, BMS-387032 and 201), olaparib (AZD-2281), ABT-888 (Veliparib), PD0332991; Rho-kinase inhibitors such as, for example AG014699, CEP 9722, MK4827, KU-0059436 (AZD2281), GSK429286; checkpoint kinase inhibitors such as, for LT-673, 3-aminobenzamide, A-966492, and AZD2461 example, AZD7762; aurora kinase inhibitors such as, for 0319 Exemplary Wnt/Hedgehog signaling pathway example, AZD1152, MLN8054 and MLN8237; PLK inhibi inhibitors include vismodegib (RG3616/GDC-0449), cyclo tors such as, for example, BI 2536, B16727 (Volasertib), pamine (11-deoxojervine) (Hedgehog pathway inhibitors) GSK46.1364, ON-01910 (Estybon); and KSP inhibitors such and XAV-939 (Wnt pathway inhibitor) as, for example, SB 743921, SB 715992 (ispinesib), 0320 Exemplary RNA polymerase inhibitors include MK-0731, AZD8477, AZ3146 and ARRY-520. amatoxins. Exemplary amatoxins include C.-amanitins, US 2013/0309192 A1 Nov. 21, 2013 37
B-amanitins, y-amanitins, e-amanitins, amanullin, amanullic 10332 X is a side chain of a natural or unnatural amino acid, amaninamide, amanin, and proamanullin. acid; 0321. In one embodiment the drug of the invention is a 0333) R, , is hydrogen or X and NR77 form a nitrogen non-natural camptothecin compound, Vinca alkaloid, kinase containing heterocyclic moiety; inhibitor (e.g., PI3 kinase inhibitor (GDC-0941 and PI-103)), 0334 Rs is NH or oxygen; MEK inhibitor, KSP inhibitor, RNA polymerse inhibitor, 0335 a is an integer from 1 to 6: PARP inhibitor, docetaxel, paclitaxel, doxorubicin, duocar 0336 c is an integer from 0 to 3: mycin, tubulysin, auristatin or a platinum compound. In spe 0337 d is an integer from 1 to 3; and cific embodiments, the drug is a derivative of SN-38, Vin 0338 f is an integer from 1 to 12. desine, vinblastine, PI-103, AZD 8330, auristatin E, 0339. Further examples of Vinca alkaloids are described in auristatin F, a duocarmycin compound, tubulysin compound, US 2010/03051.49 and US 20O2/O103136. or ARRY-52O. 0322. In another embodiment, the drug used in the inven 0340. In one embodiment the Vinca alkaloid of Formula tion is a combination of two or more drugs, such as, for (V) is a compound of Formula (VI): example, PI3 kinases and MEK inhibitors; broad spectrum cytotoxic compounds and platinum compounds; PARP inhibitors and platinum compounds; broad spectrum cyto (VI) toxic compounds and PARP inhibitors. 0323. In one embodiment, the Vinca alkaloid is a com pound of Formula (V):
(V)
wherein: (0341) Rao is hydrogen, —OH, -NH, or any of the fol lowing structures:
(1) wherein: 0324 Ra is hydrogen, —C(O)—C alkyl or —C(O)- x-rOH: chloro Substituted C- alkyl, (2) 0325 Rs is hydrogen, —CH or —CHO: OH: 0326 when R, and Rs are taken independently, Ris is hydrogen, and either R or R, is ethyl and the other is can hydroxyl: CH3 0327 when R, and Rs are taken together with the carbon (3) to which they are attached to forman oxiran ring, R is ethyl: OH: 0328 R is hydrogen, OH, amino group, alkyl amino or —C(R-oR2), R22. (4) 0329 each of Ro and Rindependently is hydrogen, C. alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated OH: Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, hydroxylated Cs cycloalkyl, polyhydroxylated Cls A CH cycloalkyl or a side chain of a natural or unnatural amino acid; (5) 0330 R is -OH, -NH, -COOH, -Rs C(O) (CH), C(H)(R)—N(H)(R), —Rs—C(O)(CH) (O CH2-CH2), N(H)(R) or —Rs (C(O)-CH(X)— NH) R-77; > --> 0331 each R- independently is hydrogen, Ce alkyl, CH Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; US 2013/0309192 A1 Nov. 21, 2013 38
-continued -continued (6) (18) O NH2: O (19) CH3 O (7) H O N x-r O y and O --- NH2: (20) O CH3 (8) O
O lus!'s > --> 1s. NH2 (9) CH, O wherein: 0342 a is an integer from 1 to 6; and O lu-Ns 0343 c is an integer from 0 to 3. 0344. In one embodiment, Rao is (10) y -CH, - O - (11) 0345. In another embodiment, non-natural camptothecin is a compound of Formula (VII):
(12)
(VII)
(13)
(14)
wherein: 0346 R is —H. —Cl, —F. —OH or alkyl; or R and Rs, may be taken together to form a five- or six-membered r1ng, (15) (0347 Rs is -H, -F, -OH, -CH, -CH=N-O-t- Butyl, —CHCHSi(CH), —Si((CH))-t-butyl, —O C (O)—Ro: 0348 Rois—NH2, —Rs Calkyl-R, 5 to 12-mem bered heterocycloalkyl, Ras Cs. heterocycloalkyl-C- alkyl-R22 or—R2s Coalkyl-C-2 aryl-C alkyl-R22. (16) (0349 R is —H, —CH N(CH), NH, or NO; 0350 R-7 is ethyl, N-methyl piperidine, cycloalkyl, —CHCH-NHCH(CH), or —N-4-methylcyclohexy lamine; 0351 Ro is —H or —C(O)—Rs C(RoR), R: (17) 0352 each of Ro and R independently is hydrogen, C. alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, hydroxylated Cls cycloalkyl, polyhydroxylated Cs cycloalkyl or a side chain of a natural or unnatural amino acid; US 2013/0309192 A1 Nov. 21, 2013 39
0353 R is -OH, -NH, -COOH, -Rs C(O) 0371 X is a side chain of a natural or unnatural amino (CH2), C(H)(R)—N(H)(R), —Rs—C(O)(CH2) (O acid; CH2-CH2), N(H)(R), or —Rs (C(O)-CH(X)— 0372) R, , is a hydrogen or X and NR-7, form a nitrogen NH) R-77; containing cyclic compound; 0354 each R- independently is hydrogen, Ce alkyl, 0373 Rs is NH or oxygen; Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 0374 c is an integer from 0 to 3: 0355 X is a side chain of a natural or unnatural amino 0375 d is an integer from 1 to 3; and acid; 0376 f is an integer from 1 to 12. 0356) R, , is a hydrogen or X and NR-7, form a nitrogen 0377. In some embodiments Ro is any one of the follow containing cyclic compound; ing structures: 0357 Rs is NH or oxygen; 0358 or R and R, when taken together with the two carbon atoms to which they attach and the third carbon atom (1) connecting the two carbon atoms form an optionally Substi tuted six-membered ring; NH-(CH2)-NH2: 0359 Rs is absent, NH or oxygen; 0360 a is an integer from 1 to 6: (2) 0361 c is an integer from 0 to 3: 0362 d is an integer from 1 to 3: 0363 f is an integer from 1 to 12: 0364 u is an integer 0 or 1: (3) 0365 w is an integer 0 or 1; and 1n-NH2: 0366 with the proviso that the compound of Formula (VII) must contain at least one of Ro and Rio NO 0367. In one embodiment the non-natural camptothecin compound of Formula (VII) is a compound of Formula (VIII) (4) or Formula (XXV): r 1-N-OH: (VIII) -
(5)
(6) O
(XXV) J.
(7)
(8)
(9) wherein Rio is —NH2, —Rs C alkyl-R, 5 to 12-mem bered heterocycloalkyl, Ras Cs. heterocycloalkyl-C alkyl-R22 or—R2s Coalkyl-C-2 aryl-C alkyl-R22. 0368 Rs is absent, NH or oxygen; 0369 R is -OH, -NH, -COOH, -Rs C(O) (CH), C(H)(R)—N(H)(R), —Rs—C(O)(CH) (O CH2-CH2), N(H)(R) or —Rs (C(O)-CH(X)— wherein: NH) R-77; 0378 a is an integer from 1 to 6: 0370 each R- independently is hydrogen, C alkyl, 0379 c is an integer from 0 to 3; and Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 0380 g is an integer from 2 to 6. US 2013/0309192 A1 Nov. 21, 2013 40
0381. In another embodiment the PI3 kinase is a com 0383 each of Ro and R independently is hydrogen, C. pound of Formula (IX): alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, hydroxylated Cs cycloalkyl, polyhydroxylated Cls
(IX) cycloalkyl or a side chain of a natural or unnatural amino acid; 0384 Rois—OH, -NHRs —N—(Rs)R.—COOH, —Rs—C(O)(CH), C(H)(R)—N(H)(R), —Rs. C (O)(CH2)A (OCH CH2), N(H)(Ras), —Rs (C(O)— CH(X)—NH), R, , or - Rs. C(O)—C(RoR) - R82 R8s; 0385 each R, independently is hydrogen, C alkyl, Caryl, Cs cycloalkyl, -COOH, or—COO Calkyl: 0386 X is a side chain of a natural or unnatural amino acid; (0387 R-7, is a hydrogen or X and NR-7, form a nitrogen containing cyclic compound; wherein 0388 Rs is NH or oxygen; 0382 Ra, is an amino group, —R C(RoR), Ro, 0389 R is absent, N—(Rs) or oxygen; —Ro Cs. heterocycloalkyl-C alkyl-Ro or 5 to 0390 Rs is hydrogen or CH: 12-membered heterocycloalkyl; 0391 R is:
O 2 O Xs O X 7 Sa NH NH R12Sa. R82 NH NH--Rs}- X4 O X6 O
0392 each R, independently is hydrogen, chloride, —CH or —OCH: 0393 R is hydrogen or —C(O)—(CH) (O—CH2— CH), NH2: 0394 X is the side chain of lysine, arginine, citrulline, alanine or glycine; 0395. X is the side chain of phenylalanine, valine, leu cine, isoleucine or tryptophan; 0396 each of X and X, is independently the side chain of glycine, alanine, serine, Valine or proline; 0397 a is an integer from 1 to 6: 0398 c is an integer from 0 to 3: 0399 d is an integer from 1 to 3: 04.00 f is an integer from 1 to 12; and 04.01 each u independently is an integer 0 or 1. 0402. In some embodiments
O Xs O X7 NH NH NHT, NH X4 O X6 O
is citrulline-valine; lysine-phenylalanine; citruline-phenyla lanine; citruline-leucine; citrulline-Valine-glycine-glycine; glycine-phenylalanine-glycine-glycine; Valine; proline; leu cine or isoleucine. US 2013/0309192 A1 Nov. 21, 2013 41
0403. In another embodiment, R is any one of the fol lowing structures: co (1) O1. NH2 (2) O
N NH2: x rol N O H N --O H H O
O1. NH2 (3) (4) O
> O O H N-1: NH2. and N NH 1st O O NH HN 1. 1. O NH2 O NH2 (5) O
Nirrri. N -----"O 3
HN 1. NH2 US 2013/0309192 A1 Nov. 21, 2013 42
04.04. In some embodiments R, is any one of the follow ing structures: (1) -- (2) (3) (4)
N --o-c- (5) J. (6) - co (7) (8) (9) (10) , CH (11) (12) -----th CH3 O (13) (14) CH3 O CH3
(15) " (C H2) O ls (16)
N 1st O US 2013/0309192 A1 Nov. 21, 2013
-continued O (17)
X -- O NH Y1 lusH N 1sH 1st and O O
es NH2 (18)
N ---, O-N-1 NH2,
HN 1. NH2 wherein: 0416 each Rs independently is hydrogen, OH, Cisalkyl, 04.05 a is an integer from 1 to 6: Cs carbocycle or O—(Cs alkyl); 0406 c is an integer from 0 to 3; and 0417 R is: 04.07 g is an integer from 2 to 6. 0408. In another embodiment the auristatin is a compound of Formula (X): O R45 R33 H O R37 CH s R39 R31 s' N X, S^ N N a. R53 R38 or Rs wherein: 10418 Rao is H. Cls alkyl, Co aryl, - X'—Cao aryl, 04.09 each of R and R independently is hydrogen or Css carbocycle, Css heterocycle, X-Cls heterocycle, Cs alkyl and at most one of R and R is hydrogen; —Cs alkylene-NH2 or (CH2)SCH 0410 R is hydrogen, Cs alkyl, Cs carbocycle, Co 0419 each X* independently is Co alkylene or Co aryl, C.s alkyl-Co-o aryl, X'-(Css carbocycle), C.s het cycloalkylene; erocycle or X'-(Css heterocycle); 0420 Raa is hydrogen or Cs alkyl; 0411 R is hydrogen, Cs alkyl, Cs carbocycle, Co aryl, X-Colo aryl.X'—(Css carbocycle), Casheterocycle 0421 Ras is X R or NH Ro: or X'—(Css heterocycle); 0422 X is O or S; 0412 R is hydrogen or methyl; 0423 R is hydrogen, OH, amino group, alkyl amino or 0413 or R and Rs, together with the carbon atom to —C(R-oR2), R22. which they attach form a carbocyclic ring having the formula 0424 R is an amino group, C alkyl amino or —C —(CRssR) - wherein each of Rss and Raindependently (R20R2), R22. is hydrogen or Cs alkyl and b is an integer from 3 to 7: 0425 each of Ro and R independently is hydrogen, C. 0414 R is hydrogen or Cs alkyl; alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated 0415 R-7 is hydrogen, Cs alkyl, Cs carbocycle, Co Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, aryl, -X'—Cao aryl, —X'—(Cs carbocycle), Cls het hydroxylated Cls cycloalkyl, polyhydroxylated Cs erocycle or - X'-(Css heterocycle); cycloalkyl or a side chain of a natural or unnatural amino acid; US 2013/0309192 A1 Nov. 21, 2013 44
0426 R is —OH, -NHR, -COOH, - Rs. C(O) 0444 wherein R is —CH or any one of the following (CH2), C(H)(R)—N(H)(R), —Rs C(O)(CH2) - Structures: (OCH CH), N(H)(Rs) or —Rs (C(O)-CH(X)— NH) R-77; 0427 each R- independently is hydrogen, Ce alkyl, Caryl, Cs cycloalkyl, -COOH, or—COO—C alkyl: (1) 0428 X is a side chain of a natural or unnatural amino OH: acid; 0429 Rz, is a hydrogen or X and NR-7, form a nitrogen x-r containing cyclic compound; (2) 0430 Rs is NH or oxygen; OH: 0431 Rs is —C(Rs.) C(Rs.) Co. aryl, —C(Rs.) C(Rs.) Cls heterocycle or -C(Rs.) C(Rs) - canCH Cascarbocycle; 0432 Rs is independently selected from H, OH, Cs (3) alkyl, Cs carbocycle. —O—Cs alkyl, —O—C(O)—R OH: and —O-R-O-C alkyl-NH2, 0433 R is an amino group, 5 to 12-membered heterocy cloalkyl, -R-2s C alkyl-R22, R2s Cs-12 heterocy cloalkyl-C alkyl-R22, —C(RoR), R22, or —R2s – C. alkyl-C-2 aryl-C alkyl-R22. 0434 Rs is absent, NH or oxygen; (4) 0435 a is an integer from 1 to 6: 0436 c is an integer from 0 to 3: OH: 0437 d is an integer from 1 to 3; and 0438 f is an integer from 1 to 12. CH3 0439. In some embodiments, in the auristatin compound of Formula (X): 0440 R is benzyl or (5) O NH2: > --> O CH3
(6) and O 0441 R4 is hydrogen; NH2: 0442. In one embodiment the auristatin of Formula (X) is O a compound of Formula (XI), Formula (XII) or Formula (XIII): CH 0443 wherein the compound of Formula (XI) is:
(XI) HC CH3 HC O CH CH3 3 NN C H3 O C H3 OCH O OCH O -S O O-R US 2013/0309192 A1 Nov. 21, 2013 45
-continued -continued (7) (14) O CH3 O
O 1- NH2: O NH2:
(8) CH3 O NH2: CH3 O O *--- H (15) CH, O
O lu-Ns (16) O O CH, O (10) I-3- no ul-1-2 N-1s1-12YNH A-N-1- NH2:2 (17)
(11) (18) O CH lul C(H)(CH3)-(CH2)NH2: O NH2: (19) O
N (12) x-r O yH and CH, O CH (20) O CH3 O NH2: car-nO CH3: NH2 (13) O CH 3 wherein:
Olul NH2: 0445 a is an integer from 1 to 6; and 0446 c is an integer from 0 to 3: 0447 wherein the compound of Formula (XII) is:
(XII) H3C CH3 H3C O CH CH3 3 NN CH, O -S CH, OCH O OCH O O NH R40 US 2013/0309192 A1 Nov. 21, 2013 46
0448 wherein Rao is hydrogen, OH, -NH2 or any of -continued the following structures: (10) CH, O
(1) y - - - >n->OH: (11) (2)
(12) CH3 (3) >n-" (13) (4) OH: (14) CH3 (5) O NH2: O (15) CH3
(6) (16) O NH2: O CH (17)
(7) O (18)
O --- NH2: (19) O H N O (8) O x-r y and (20) O lu-Ns O CH3 > --> 1s. NH2 (9) CH3 O wherein: O --- NH2: 0449) a is an integer from 1 to 6; and 0450 c is an integer from 0 to 3. US 2013/0309192 A1 Nov. 21, 2013 47
0451 wherein the compound of Formula (XIII) is:
(XIII) HC CH3 HC Y-Rs O CH CH3 O 3 NN CH, O HC 1Ne,E CH, OCH O OCH O CH
0452 wherein R is an amino group, 5 to 12-membered C- alkyl-R22, R2s Cs-12 heterocycloalkyl-C alkyl-R22 heterocycloalkyl, —Rs Calkyl-R, R2s Cs-hetero or R2s Co alkyl-C-2 aryl-C alkyl-R22. cycloalkyl-C alkyl-R22. —R2s C(RoR), R22, or —R2s C1-alkyl-C-2 aryl-C- alkyl-R22. 0466 Rs is absent, NH or oxygen; 0453 each of Ro and R independently is hydrogen, C. 0467 R is —OH, -NHR, —COOH, - Rs. C(O) alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated (CH), C(H)(R)—N(H)(R), RC(O)(CE) - Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, (OCH2-CH2), N(H)(R) or —Rs—(C(O)—CH(X)— hydroxylated Cs cycloalkyl, polyhydroxylated Cls NH) R-77; cycloalkyl or a side chain of a natural or unnatural amino acid; 0468 each R, independently is hydrogen, C alkyl, 0454 R is —OH, -NHR, -COOH, - Rs. C(O) Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 10469 X is a side chain of a natural or unnatural amino (OCH(CH). CH)(R)N(H)(R),CH), N(H)(Rs) or Rs RC(O)(CH),(C(O) CH(X)— acid; NH) R-77; 0470 R7, is a hydrogen or X and NR-7, form a nitrogen 0455 each R- independently is hydrogen, Ce alkyl, containing cyclic compound; Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 0456 X is a side chain of a natural or unnatural amino 0471) Rs is —NH or oxygen; acid; 0472. c is an integer from 0 to 3: 0457 R-7, is a hydrogen or X and NR-7, form a nitrogen 0473 d is an integer from 1 to 3; and containing cyclic compound; 0474 f is an integer from 1 to 12. 0458 Rs is NH or oxygen; 0475. In yet another embodiment, R is any one of the 0459 Rs is absent, NH or oxygen; following structures: 0460 a is an integer from 1 to 6: 0461 c is an integer from 0 to 3: 0462 d is an integer from 1 to 3; and (1) 0463 f is an integer from 1 to 12. 0464) In one embodiment, in Formula (XII), Rao is
(2)
(3)
O NH2, or. cr O (4) CH
car." (5) CH
0465. In one embodiment in the compound of Formula (XIII), R is —NH 5 membered heterocycloalkyl, —Rs— US 2013/0309192 A1 Nov. 21, 2013 48
-continued 0491 d is an integer from 1 to 3; and (6) 0492 f is an integer from 1 to 12. 0493. Further examples of the MEK inhibitor are dis closed in U.S. Pat. No. 7,517,994 B2. x. 0494. In some embodiments R is —C(O)—(CH)— (7) NH, or—C(O)—C(H)(CH)–(CH), NH; in which a is an integer from 1 to 6; and c is an integer from 0 to 3. -- (CH2)-NH2: 0495. In another embodiment, the duocarmycin com pound is a compound of Formula (XV): (8) - one-class and (XV) , (9) wherein: 0476 a is an integer from 1 to 6: 0477 c is an integer from 0 to 3; and 0478 g is an integer from 2 to 6. 0479. In one embodiment, the MEK inhibitor is a com pound of Formula (XIV): wherein: 0496 Ra, is as defined herein; (XIV) H 0497 Ras is hydrogen, —COOC alkyl, -COOH, O N OR43 F no-1S-1 —NH2 or —CH: 0498 R is C1, Br or -OH: 0499 Rso is hydrogen, —OCH,
0480 wherein R is H or —Ra Raz; 0481 each of Ro and Rindependently is hydrogen, C. alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, hydroxylated Cls cycloalkyl, polyhydroxylated Cls cycloalkyl or a side chain of a natural or unnatural amino acid; 0482 R is —OH, -NH2, —COOH, - Rs. C(O) (CH), C(H)(R)—N(H)(R), —Rs—C(O)(CH) - (OCH CH), N(H)(Rs) or —Rs (C(O)-CH(X)— NH), R77; 0483 each R- independently is hydrogen, Ce alkyl, Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 0484 X is a side chain of a natural or unnatural amino 0500 each of Rs and Rs independently is hydrogen or acid; 0485 R7, is a hydrogen or X and NR-7, form a nitrogen —OCH; and containing cyclic compound; 0501) ring AA is either a phenyl or pyrrolyl ring. 0486 Rs is NH or oxygen; 0502. Further examples of duocarmycin compounds are 0487 R is —C(O)— —C(O)—O , C(O) NH-, or absent; disclosed in U.S. Pat. No. 7,553,816. 0488 R, is as defined herein; 0503. In one embodiment the duocarmycin compound of 0489 a is an integer from 1 to 6: Formula (XV) is a compound of Formula (XVI), (XVII), 0490 c is an integer from 0 to 3: (XVIII) or (XIX): US 2013/0309192 A1 Nov. 21, 2013 49
(XVI) (XVII) OCH OCH R49 C HCO O | OCH | OCH N N 7 NH OCH NH OCH O O N H s' s' O O
(XVIII) C H3C
s
C N NH N O H3C I N O OH N N / H O N H s" O
(XIX)
H / s C N O O H3C I N N 7 H O N H s" O wherein: 0504 R is C1, Br or -OH; and 0505) R, is as defined herein. 0506. In another embodiment, the duocarmycin com pound is a duocarmycin SA compound of Formula (XX): U.S. Pat. No. 5,101,038; or (XXI): US 2013/0309192 A1 NOV 21, 2013 50
-continued (XX) (2) OCH
OCH CH3 (3) O N / H OCH3 R42-O O >n-" N H (4) "S R47 OH: O (XXI) O (5) H3C O-R
r Br HN OCH (6) HCN NCOO N OCH \-/ 4N N OCH (7) wherein: 0507 R is C alkyl amino or —C(RoR), R: 0508 each of Ro and R independently is hydrogen, C. alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated Caryl, 5 to 12-membered heterocycle, C.s cycloalkyl, (8) hydroxylated Cls cycloalkyl, polyhydroxylated Cls cycloalkyl or a side chain of a natural or unnatural amino acid; 05.09 R is -OH, -NH, -COOH, -Rs C(O) (CH), C(H)(R)—N(H)(R), —Rs—C(O)(CH) (O (9) CH2-CH2), N(H)(R), or —Rs (C(O)-CH(X)— NH) R-77; 0510 each R- independently is hydrogen, Ce alkyl, Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 0511 X is a side chain of a natural or unnatural amino (10) acid; 0512) R, , is a hydrogen or X and NR-7, form a nitrogen containing cyclic compound; 0513 Rs is —NH or oxygen; (11) 0514 a is an integer from 1 to 6: 0515 c is an integer from 0 to 3: 0516 d is an integer from 1 to 3; and 0517 f is an integer from 1 to 12. (12) 0518. In some embodiments, R is any one of the follow ing structures:
(13) > -->OH: (1) US 2013/0309192 A1 Nov. 21, 2013
-continued 0529 R is C alkyl, —CHs or —CH-phenyl: (14) 0530 R is C alkyl; CH, O 0531 R is hydrogen, OH, O C alkyl or O C(O)— NH2: Calkyl, O 0532 R is hydrogen, OH, O C alkyl, O C(O)— CH3 C. alkyl, halogen or C. alkyl, (15) 0533 e is an integer between 1 and 3 inclusive: CH, O 0534 R is: N O R71
(16) R. O
1-3O1-4N-). 1-2 1-12YNH. R73 O (17)
R68 R69 R71 (18) R72:
R73 (19) O R45 H N O O ; and
(20) wherein: O CH3 0535 Rs is hydrogen or C-C alkyl: 0536 R. is COR, C(O)-Rs, CONHNH, OH, NH, SH or optionally substituted alkyl, cycloalkyl, heteroalkyl or >n-> 1. heterocycloalkyl group; NH2 0537 Rao is an optionally substituted alkyl (i.e. C. alkyl amine), heteroalkyl or heterocycloalkyl group; wherein: 0538 each of R, and R independently is hydrogen, 0519 a is an integer from 1 to 6; and halo. —NO. —CN, -NHRC alkyl, haloalkyl, alkoxy, 0520 c is an integer from 0 to 3. and haloalkoxy; 0521. In another embodiment the tubulysin is a compound 0539 R is hydrogen, OR, alkoxy, halogen, NHR, of Formula (XXII): —O—C(O)—R7, NO. —CN, Co aryl, Ce alkyl, amino or dialkylamino; (XXII) (0540 R is hydrogen, —CHO,-CO) Calkyl, OH, amino group, alkyl amino or -C(RoR), R22. ) 8 H O R6 1 R62 R63 O 0541 R is H or —Rae R7; N 0542. Rae is —C(O)— —C(O)—O—, C(O) NH-, N-R or absent; Nk O R59 R60 --S 7 H 0543. Ra, is as defined herein; (0544 Ris is X Rs or NH Ro: 0522 wherein: (0545 X is O or S; 0523 Rs, is C alkyl or —C(O)Rss; 0546 Rio is hydrogen, OH, amino group, alkyl amino or 0524 Rss is C alkyl, CF or Co aryl; —C(R-oR2), R22. 0525 Rs is C alkyl: 0547 R-7s is a hydrogen, an amino group, Ce alkylamino 0526 Rao is hydrogen, Ce alkyl, C-, alkenyl, -CH2 or —C(RoR2), R22. phenyl, CHORs or CHOCOR; 0548 each of Ro and R independently is hydrogen, C. 0527. Resis hydrogen, Ce alkyl, C-7 alkenyl, Co aryl alkyl, Co aryl, hydroxylated Co aryl, polyhydroxylated or C(O)R7; Co aryl, 5 to 12-membered heterocycle, C.s cycloalkyl, 0528 R, is C alkyl, C- alkenyl, Co aryl or het hydroxylated Cls cycloalkyl, polyhydroxylated Cs eroaryl; cycloalkyl or a side chain of a natural or unnatural amino acid; US 2013/0309192 A1 Nov. 21, 2013 52
(0549. R. is -OH, -NH, -COOH, -Rs C(O) -continued (CH2), C(H)(R)—N(H)(R), —Rs—C(O)(CH) (5) (O CH2 -CH2), N(H)(R), or —Rs—(C(O)—CH O (X) NH), R77; NH2: 0550 each R- independently is hydrogen, C alkyl, O Co aryl, Css cycloalkyl, -COOH, or—COO Cealkyl; 0551 X is a side chain of a natural or unnatural amino CH3 (6) acid; O 0552) R, , is a hydrogen or X and NR-7, form a nitrogen containing cyclic compound; NH2: 0553 Rs is NH or oxygen; O 0554 Ra, is as defined herein; CH3 0555 a is an integer from 1 to 6: (7) 0556 c is an integer from 0 to 3: O 0557 d is an integer from 1 to 3: 0558 f is an integer from 1 to 12; and O --Ns 0559 with the proviso that when R is C(O) X-Rs or (8) C(O) NH Rio, one or both of R, and R7 are —NHR, O and R is OR —NHR, or—O—C(O)—Raz, at least one of Rio, Ras, R-7 and R-7s cannot be hydrogen. 0560. In some embodiments in the compound of Formula O lu-Ns (XXII): (9) 0561 Rs, is CH: CH, O 0562 Rs is sec-butyl: 0563 Reo is hydrogen, methyl, ethyl, propyl, iso-propylor O lu-Ns iso-butyl: (10) 0564 R is iso-propyl, CH, O 0565 R is hydrogen; 0566 R is hydrogen, OH, - O CH7, O—C(O)— O lus!'s CH: (11) 0567 Rs is hydrogen or —CH: O CH3 0568 R is COH, COR, or C(O)—R-7s; 0569 Rao is C alkyl amine: 0570 each of R, and R, independently is hydrogen; > --> 0571 R is hydrogen, —OR, OH, F, —CH or (12) –OCH: CH, O CH3 0572 Rs is OH, -ORs or - NHR: 0573 e is the integer 2: 0574 Rao is hydrogen, —OH, -NH, or any of the fol A-s-s-s (13) lowing structures: O CH3
(1) ->-Nulls NH2: (14) >n-> OH: CH, O NH2: (2) O OH: CH3 can (15) CH3 CH3 O (3) OH: N Xu O y (4) (16) O OH: N-). * -- -Yo 1-2 1-12YNH CH US 2013/0309192 A1 Nov. 21, 2013 53
-continued -continued (17) (8) O
O lus!'s (18) (9) CH, O
O lu-Ns (19) (10) CH, O O and O lu-Ns (20) (11) O CH3 O CH > --> 1. > --> NH2 (12) CH, O CH wherein: (0575 a is an integer from 1 to 6: 0576 c is an integer from 0 to 3: (13) 0577 R.7s is any one of the following structures: O CH
(1) <-->--- NH: (14) CH, O NH2: (2) O CH (15) CH3 CH, O (3) N can- OH: O (4) (16) O OH: -Yo 1-2 'N1).1-12YNH. (5) (17)
(CH2)-NH2; and
(18) (6)
wherein: (7) 0578 a is an integer from 1 to 6; and (0579 c is an integer from 0 to 3: 0580 R is hydrogen, —C(O) (CH), NH, O US 2013/0309192 A1 Nov. 21, 2013 54 wherein: 0581 a is an integer from 1 to 6: 0582 c is an integer from 0 to 3; and 0583 Ra, is any one of the following structures: (1) ---it-on (2) (3) OH: (4) xr^- s --o-c- (5) x - (6) - co (7) - one-class (8) (9) (10) --t -(CH2)-OH:
(11) (12) -----th ------t (13) (14) -- CH3 O ---a--t N (15) --to-lt NH2 (16) N-1 H s N N 1st O US 2013/0309192 A1 Nov. 21, 2013 55
-continued (17) X N N-1 n1nols O N-1 o H H N 1s 1st O O O
HN es NH2 (18) x-r's O N-1 o N H 's N ---, O -N1"3 O esHN wherein: 0587 with the proviso that if R is —OH, then Rs cannot 0584 a is an integer from 1 to 6: be hydrogen; if R is COOH then R, must be —OR or —O—C(O)—R7. 0585 c is an integer from 0 to 3; and 0588. In some embodiments, the tubulysin of Formula 0586 g is an integer from 2 to 6; (XXII) is a compound of Formula (XXIII) or (XXIV):
(XXIII) R76
OCH, O
(XXIIIa) R76
ClCH, O O-Rs: US 2013/0309192 A1 Nov. 21, 2013 56
-continued (XXIIIb)
O
1 i N
(XXIV)
OJCH l, O s shi,
wherein: 0594. In some embodiments R is: 0589 R2 is hydrogen, OH, OCH. F. —OR or —O C (1) (see0590 a hereinere1n R-7s., R.7s.,R-7s, Rio,Ro RR47 andd RR4 are as defineddeline NH-(CH3)-NH2: 0591 with the proviso that if R is -OH, OCH or F, then R7s and R cannot be hydrogen. (2) 0592. In one embodiment, R, is NH-(CH2)-OH:
HN (3) NH2: 0593. In another embodiment, the KSP inhibitor com- r 1N1 pound is a compound of Formula (XXVI): x
(XXVI)
(4) OH xr^- s
F (5) N \ O-(CH2)-NH2: wherein Rio is as defined herein. US 2013/0309192 A1 Nov. 21, 2013 57
-continued -continued (6) (XXVIII)
x. O N K 2 (7) S N -- (CH2)-NH2 or O
NHR11 - A coco -(CH2)NH2: (8)
(XXIX) (9) F N-6 N HN 2Y-k O F S
RHN wherein: 0595 a is an integer from 1 to 6: 0596 c is an integer from 0 to 3; and 0597 g is an integer from 2 to 6. 0598. In another embodiment the KSP inhibitor com pound is a compound of Formula (XXVII), (XXVIII) or (XXIX): wherein: 0599 R is as defined herein. (XXVII) 0600. One skilled in the art of therapeutic agents will readily understand that each of the therapeutic agents described herein can be modified in such a manner that the resulting compound still retains the specificity and/or activity of the original compound. The skilled artisan will also under stand that many of these compounds can be used in place of the therapeutic agents described herein. Thus, the therapeutic C N agents of the present invention include analogues and deriva tives of the compounds described herein. O 0601 Table B below provides more examples of the thera peutic agents and derivatives thereofsuitable for conjugation to form the polymer-drug-protein conjugates or polymer drug scaffolds of the invention. Spectral data of certain com 2. NHR11 pounds are also provided (ND in the table means “not deter mined'). These examples may also be the active form of the drug when it is released from the conjugates invitro or in vivo. US 2013/0309192 A1 Nov. 21, 2013 58
TABLE B
(VI)
Ref# Rao
Ex22
Ex 23 Sry OH
(IX)
Ex 24 NH ND Cr- 2
Ex 2S H ND
US 2013/0309192 A1 Nov. 21, 2013 60
TABLE B-continued
ExSO O 874.5 NH2 O
- OH 788 Ex 61 xCH3 8O3.4 EX62 Xu,CH3 8O3.4 A-s-s-CH3 O 874.4 CH3 A CH3 1O 874.4 CH3
90O.S
H2Nii, O
O US 2013/0309192 A1 Nov. 21, 2013 61
TABLE B-continued (XIII)
H3C CH3 H3C X- R29 O CH CH3 O 3 NN CH, O HC1E NCH, CH, OCH O OCH O CH3
mz
903.2 Sk 8O3.1
O 790 NH2
O ND
829.1
802
(XIV)
Ex36 ND US 2013/0309192 A1 Nov. 21, 2013 62
TABLE B-continued
E O O 644.9
O
(XVII) OCH C
OCH ... I N OCH O s' O
R47 mz
553.1
S38.1
564.1
566.1
568.1
ND US 2013/0309192 A1 Nov. 21, 2013 63
TABLE B-continued
ND
667.2
622.2
ND
ND
US 2013/0309192 A1 Nov. 21, 2013 65
TABLE B-continued ~~ - OH ND
---O
- OH 871.4
19 NH2
O
Ex 42 ND
Ex 43 ND
(XXX)
Ex 46
HN
Protein-Based Recognition Molecules (PBRMs) affinity and avidity. In some embodiments, the protein-based recognition molecule targets the modified polymer to tissues 0602. The protein-based recognition molecule directs the other than the liver. In other embodiments the protein-based drug-polymer carrier conjugates to specific tissues, cells, or recognition molecule targets the modified polymer to a spe locations in a cell. The protein-based recognition molecule cific tissue such as the liver, kidney, lung or pancreas. The can direct the modified polymer in culture or in a whole protein-based recognition molecule can target the modified organism, or both. In each case, the protein-based recognition polymerto a target cell Such as a cancer cell. Such as a receptor molecule has a ligand that is present on the cell Surface of the expressed on a cell Such as a cancer cell, a matrix tissue, or a targeted cell(s) to which it binds with an effective specificity, protein associated with cancer Such as tumor antigen. Alter US 2013/0309192 A1 Nov. 21, 2013 66 natively, cells comprising the tumor vasculature may be tar alemtuzumab (CAMPATH), altumomab, amatuximab, ana geted. Protein-based recognition molecules can direct the tumomab, anrukinZumab, apolizumab, arcitumomab (CEA polymer to specific types of cells such as specific targeting to SCAN), aselizumab, atlizumab (tocilizumab, Actemra, hepatocytes in the liver as opposed to Kupffer cells. In other RoActemra), atorolimumab, bapineuZumab, basiliximab cases, protein-based recognition molecules can direct the (Simulect), bavituximab, bectumomab (LYMPHOSCAN), polymer to cells of the reticular endothelial or lymphatic belimumab (BENLYSTA), benralizumab, bertilimumab, system, or to professional phagocytic cells Such as macroph besilesomab (SCINITIMUN), bevacizumab (AVASTIN), ages or eosinophils. (In Such cases the polymer itself might biciromab (FIBRISCINT), bivatuzumab, blinatumomab, also be an effective delivery system, without the need for brentuximab, briakinumab, canakinumab (ILARIS), cantu specific targeting). Zumab, capiromab, catumaxomab (REMOVAB), CC49, 0603 Instill other embodiments, the protein based recog cedelizumab, certolizumab, cetuximab (ERBITUX), citatu nition molecule can target the modified polymer to a location Zumab, cixutumumab, clenoliximab, clivatuZumab, conatu within the cell. Such as the nucleus, the cytoplasm, or the mumab, CR6261, dacetuzumab, daclizumab (ZENAPAX). endoSome, for example. In specific embodiments, the protein daratumumab, denosumab (PROLIA), detumomab, dorlimo based recognition molecule can enhance cellular binding to mab, dorlixizumab, ecromeximab, eculizumab (SOLIRIS), receptors, or cytoplasmic transport to the nucleus and nuclear edobacomab, edrecolomab (PANOREX), efalizumab (RAP entry or release from endosomes or other intracellular TIVA), efungumab (MYCOGRAB), elotuzumab, elsilimo vesicles. mab, enlimomab, epitumomab, epratuZumab, erlizumab, 0604. In specific embodiments the protein based recogni ertumaxomab (REXOMUN), etaracizumab (ABEGRIN), tion molecules include antibodies, proteins and peptides or exbivirumab, fanolesomab (NEUTROSPEC), faralimomab, peptide mimics. farletuZumab, felvizumab, fezakinumab, figitumumab, fon 0605 Exemplary antibodies or antibodies derived from tolizumab (HuZAF), foravirumab, fresolimumab, galiximab, Fab, Fab2, scFv or camel antibody heavy-chain fragments gantenerumab, gavilimomab, gemtuzumab girentuximab, specific to the cell Surface markers, include, but are not lim glembatumumab, golimumab (SIMPONI), gomiliximab, ited to, 5T4, AOC3, C242, CA-125, CCL11, CCR5, CD2, ibalizumab, ibritumomab, igovomab (INDIMACIS-125), CD3, CD4, CDS, CD15, CD18, CD19, CD20, CD22, CD23, imciromab (MYOSCINT), infliximab (REMICADE), intetu CD25, CD28, CD30, CD31, CD33, CD37, CD38, CD40, mumab, inolimomab, inotuZumab, ipilimumab, iratumumab, CD41, CD44, CD51, CD52, CD54, CD56, CD62E, CD62P, keliximab, labetuzumab (CEA-CIDE), lebrikizumab, CD62L, CD70, CD74, CD80, CD125, CD138, CD141, lemalesomab, lerdelimumab, lexatumumab, libivirumab, lin CD147, CD152, CD 154, CD326, CEA, clumping factor, tuZumab, lucatumumab, lumiliximab, mapatumumab, masli CTLA-4, EGFR, ErbB2, ErbB3, EpCAM, folate receptor, momab, matuzumab, mepolizumab (BOSATRIA), mete FAP, GD2, GD3, GPNMB, HGF, HER2, ICAM, IGF-1 limumab, milatuZumab, minretumomab, mitumomab, receptor, VEGFR1, EphA2, TRPV1, CFTR, gpNMB, CA9, morolimumab, motavizumab (NUMAX), muromonab-CD3 Cripto. ACE, APP, adrenergic receptor-beta2, Claudine 3, (ORTHOCLONEOKT3), nacolomab, naptumomab, natali Mesothelin, IL-2 receptor, IL-4 receptor, IL-13 receptor, inte Zumab (TYSABRI), nebacumab, necitumumab, nerelimo grins (including C-4, Clfs. Clfs. Clfg. Clifa, Cla?'. Cla?, mab, nimotuzumab (THERACIM), nofetumomab, ocreli Os3, CB, CB intergins), IFN-O, IFN-Y, IgE. IgE, IGF-1 Zumab, odulimomab, ofatumumab (ARZERRA), receptor, IL-1, IL-12, IL-23, IL-13, IL-22, IL-4, IL-5, IL-6, olaratumab, omalizumab (XOLAIR), ontecizumab, oportu interferon receptor, ITGB2 (CD18), LFA-1 (CD11a), L-se Zumab, oregovomab (OVAREX), otelixizumab, pagibaxi lectin (CD62L), mucin, MUC1, myostatin, NCA-90, NGF, mab, palivizumab (SYNAGIS), panitumumab (VECTIBIX), PDGFRC, phosphatidylserine, prostatic carcinoma cell, panobacumab, pascolizumab, pemtumomab (THERAGYN), Pseudomonas aeruginosa, rabies, RANKL, respiratory syn pertuzumab (OMNITARG), pexelizumab, pintumomab, pril cytial virus, Rhesus factor, SLAMF7, sphingosine-1-phos iximab, pritumumab, PRO 140, rafivirumab, ramucirumab, phate, TAG-72, T-cell receptor, tenascin C, TGF-1, TGF-32, ranibizumab (LUCENTIS), raxibacumab, regavirumab, TGF-B, TNF-ct, TRAIL-R1, TRAIL-R2, tumor antigen reslizumab, rilotumumab, rituximab (RITUXAN), robatu CTA A16.88, VEGF-A, VEGFR2, vimentin, and the like. mumab, rontalizumab, rovelizumab (LEUKARREST), rupli 0606. In one embodiment the antibodies or antibody Zumab (ANTOVA), satumomab pendetide, Sevirumab, sibro derived from Fab, Fab2, scFv or camel antibody heavy-chain tuZumab, Sifalimumab, siltuximab, siplizumab, fragments specific to the cell Surface markers include SolaneZumab, Sonepcizumab, Sontuzumab, Stamulumab, CA-125, C242, CD3, CD19, CD22, CD25, CD30, CD31, sulesomab (LEUKOSCAN), tacatuzumab (AFP-CIDE), tet CD33, CD37, CD40, CD44, CD51, CD54, CD56, CD62E, raxetan, tadocizumab, talizumab, taneZumab, taplitumomab CD62P, CD62L, CD70, CD138, CD141, CD326, CEA, paptox, tefibazumab (AUREXIS), tellimomab, tenatumomab, CTLA-4, EGFR, ErbB2, ErbB3, FAP, folate receptor, IGF-1 teneliximab, teplizumab, TGN1412, ticilimumab (tremeli receptor, GD3, GPNMB, HGF, HER2, VEGF-A, VEGFR2, mumab), tigatuzumab, TNX-650, tocilizumab (atlizumab, VEGFR1, EphA2, EpCAM, 5T4, TAG-72, tenascin C, ACTEMRA), toralizumab, tositumomab (BEXXAR), trastu TRPV1, CFTR, gpNMB,CA9, Cripto, ACE, APP, PDGFRC, Zumab (HERCEPTIN), tremelimumab, tucotuzumab, phosphatidylserine, prostatic carcinoma cells, adrenergic tuvirumab, urtoxazumab, ustekinumab (STELERA), receptor-beta2, Claudine 3, mucin, MUC1, Mesothelin, IL-2 vapaliximab, vedolizumab, VeltuZumab, Vepalimomab, receptor, IL-4 receptor, IL-13 receptor and integrins (includ visilizumab (NUVION), volociximab (HUMASPECT), ing Clfs. Clfs. Clfg. Clif4: Cla?s, Os?i, Co?sa intergins), tena votumumab, Zalutumumab (HuMEX-EGFr), Zanolimumab scin C, TRAIL-R2 and vimentin. (HuMAX-CD4), Ziralimumab and Zolimomab. 0607 Exemplary antibodies include 3F8, abagovomab, 0608. In some embodiments the antibodies are directed to abciximab (REOPRO), adalimumab (HUMIRA), adecatu cell surface markers for 5T4, CA-125, CEA, CD3, CD19, mumab, afelimomab, afutuzumab, alacizumab, ALD518, CD2O, CD22, CD30, CD33, CD40, CD44, CD51, CTLA-4, US 2013/0309192 A1 Nov. 21, 2013 67
EpCAM, HER2, EGFR, FAP, folate receptor, HGF, integrin derived peptides, ApoA protein peptides, Somatostatin recep O?3 integrin Os3, IGF-1 receptor, GD3, GPNMB, mucin, tor targeting peptides, chlorotoxin derived peptides, and MUC1, phosphatidylserine, prostatic carcinoma cells, bombesin. PDGFR C, TAG-72, tenascin C, TRAIL-R2, VEGF-A and 0611. In specific embodiments the peptides or peptide VEGFR2. In this embodiment the antibodies are abago mimics are LHRH receptor targeting peptides and ErbB2 Vomab, adecatumumab, alacizumab, altumomab, anatumo (HER2) receptor targeting peptides 0612 Exemplary proteins comprise insulin, transferrin, mab, arcitumomab, bavituximab, bevacizumab (AVASTIN), fibrinogen-gamma fragment, thrombospondin, claudin, apo bivatuZumab, blinatumomab, brentuximab, cantuzumab, lipoprotein E, Affibody molecules such as, for example, catumaxomab, capromab, cetuximab, citatuZumab, clivatu ABY-025, Ankyrin repeat proteins, ankyrin-like repeats pro Zumab, conatumumab, dacetuZumab, edrecolomab, epratu teins and synthetic peptides. Zumab, ertumaXomab, etaracizumab, farletuZumab, figitu 0613. In some embodiments of the invention the protein mumab, gemtuzumab, glembatumumab, ibritumomab, drug polymer conjugates comprise broad spectrum cytotox igovomab, intetumumab, inotuzumab, labetuZumab, lexatu ins in combination with cell surface markers for HER2 such mumab, lintuZumab, lucatumumab, matuZumab, mitumo as pertuzumab or trastuzumab; for EGFR such as cetuximab: mab, naptumomab estafenatox, necitumumab, oportuZumab, for CEA such as labetuzumab; for CD20 such as rituximab, oregovomab, panitumumab, pemtumomab, pertuzumab, pri for VEGF-A such as bevacizumab; or for CD-22 such as tumumab, rituximab (RITUXAN), rilotumumab, robatu epratuZumab or VeltuZumab. mumab, Satumomab, sibrotuZumab, taplitumomab, tenatu 0.614. In other embodiments of the invention the protein momab, tenatumomab, ticilimumab (tremelimumab), drug-polymer conjugates or protein-polymer conjugates used tigatuzumab, trastuzumab (HERCEPTIN), to situmomab, in the invention comprise combinations of two or more pro tremelimumab, tucotuZumab celmoleukin, Volocliximab and tein based recognition molecules. Such as, for example, com Zalutumumab. bination of bispecific antibodies directed to the EGF receptor (EGFR) on tumor cells and to CD3 and CD28 on T cells; 0609. In specific embodiments the antibodies directed to combination of antibodies or antibody derived from Fab, cell surface markers for HER2 are pertuzumab or trastu Fab2, ScPV or camel antibody heavy-chain fragments and Zumab and for EGFR the antibody is cetuximab and for CD20 peptides or peptide mimetics; combination of antibodies or the antibody is rituximab and for VEGF-A is bevacizumab antibody derived from Fab, Fab2, scFv or camel antibody and for CD-22 the antibody is epratuzumab or veltuzumab heavy-chain fragments and proteins; combination of two and for CEA the antibody is labetuzumab. bispecificantibodies such as CD3X CD19 plus CD28X CD22 0610 Exemplary peptides or peptide mimics include inte bispecific antibodies. grin targeting peptides (RGD peptides), LHRH receptor tar 0615 Table C below provides more examples of the geting peptides, ErbB2 (HER2) receptor targeting peptides, PBRM described hereof, which are suitable for conjugation prostate specific membrane bound antigen (PSMA) targeting to form the polymer-drug-protein conjugates or polymer peptides, lipoprotein receptor LRP1 targeting, ApoE protein PBRM Scaffolds of the invention. TABLE C
Ref# PBRM
Ex3 O
N N H O NS TRASTUZUMAB
O
Ex 4 O O
W N-n O in- -TRASTUZUMAB 2 N US 2013/0309192 A1 Nov. 21, 2013 68
TABLE C-continued
Ref# PBRM
Ex S3 O
N N O YRITUXIMAB
O
Ex 60 TRASTUZUMAB-Fab-SH
O O / H N N 1 TRASTUZUMAB-Fab N- (-1\o 2 NH O O
OH
O H O O O O N O H H N N NH2 ul N N N N N H iNH O O N-NHN1 to l NH HN -l. HN NH
2 SH
US 2013/0309192 A1 Nov. 21, 2013 70
Linkers (LP and L) (0623) In some embodiments the linker L’or L is cleaved 0616. As described above, the drug or PBRM is connected by esterases. Only certain esters can be cleaved by esterases to the polymeric carrier via a linker LP or L. In some present inside or outside cells. Esters are formed by the con embodiments, the linker is biocleavable/biodegradable under densation of a carboxylic acid and an alcohol. Simple esters intracellular conditions, such that the cleavage of the linker are esters produced with simple alcohols, such as aliphatic releases the drug or PBRM from the polymer unit in the alcohols, and Small cyclic and Small aromatic alcohols. intracellular environment. 0624. In yet other embodiments, the linker LP or L is not 0617. A linker is any chemical moiety that is capable of biocleavable and the drug is released by antibody degrada linking a drug or a PBRM to a polymer backbone through tion. See, for example, U.S. Pat. No. 7,498,298, which is chemical bonds such that the drug or PBRM and the polymer incorporated by reference herein in its entirety and for all are chemically coupled (e.g., covalently bonded) to each purposes. other. In some embodiments, the linker comprises a biode 0625 Typically, the linker LP or L is not substantially gradable linker moiety (e.g., a biodegradable bond Such as an sensitive to the extracellular environment. As used herein, ester or amide bond). “not substantially sensitive to the extracellular environment.” 0618. In other embodiments, the linker LP or L is biode in the context of a linker, means that no more than about 20%, gradable under mild conditions, i.e., conditions within a cell typically no more than about 15%, more typically no more under which the activity of the drug is not affected. Examples than about 10%, and even more typically no more than about of suitable biodegradable linker moiety include disulfide 5%, no more than about 3%, or no more than about 1% of the linkers, acid labile linkers, photolabile linkers, peptidase linkers, in a sample of Polymer Drug Conjugate, are cleaved labile linkers, and esterase labile linkers. when the Polymer Drug Conjugate presents in an extracellu 0619. In some embodiments, the linker LP or L is bio lar environment (e.g., in plasma) for 24 hours. Whether a cleavable under reducing conditions (e.g., a disulfide linker). linker is not substantially sensitive to the extracellular envi In this embodiment the drug or PBRM moiety is linked to the ronment can be determined, for example, by incubating the polymer through a disulfide bond. The linker molecule com Polymer Drug Conjugate with plasma for a predetermined prises a reactive chemical group that can react with the drug. time period (e.g., 2, 4, 8, 16, or 24hours) and then quantitating Preferred reactive chemical groups for reaction with the drug the amount of free drug present in the plasma. or PBRM moiety are N-succinimidyl esters and N-sulfosuc (0626. In embodiments, the linker LP has the structure: cinimidyl esters. Additionally the linker molecule comprises R. C(-O) X-MP1-Y-MP2-ZD-MP-Q-MP-, with a reactive chemical group, preferably a dithiopyridyl group R' connected to an oxygenatom of the polymeric carrier and that can react with the drug to form a disulfide bond. In some MP connected to the drug molecule to be delivered. embodiments the linker molecules include, for example, 0627. In embodiments, the linker L has the structure: Lif N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP), is a linker having the structure: R' C(=O) X-M'- N-succinimidyl 4-(2-pyridyldithio)butanoate (SPDB), Y-M-Z-M-Q-M-, with R', connected to an oxygen N-succinimidyl 4-(2-pyridyldithio)pentanoate (SPP), N-suc atom of the polymeric carrier and M connected to the cinimidyl-5-acetylthioacetate (SATA) and N-succinimidyl PBRM. oxycarbonyl-alpha-methyl-alpha-(2-pyridyl-dithio)toluene (0628. For example, each of R'' and R' independently is or 2,5-dioxopyrrolidin-1-yl 4-(1-(pyridin-2-yldisulfanyl) absent, alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, het ethyl)benzoate (SMPT). eroalkenyl, heteroalkynyl, heterocycloalkyl, aryl, or het 0620. In other embodiments, the biocleavable linker LP or eroaryl. L is pH-sensitive, i.e., sensitive to hydrolysis at certain pH 0629. For example, each of R'' and R' independently is values. Typically, the pH-sensitive linker is hydrolysable absent, alkyl, cycloalkyl, heteroalkyl, or heterocycloalkyl. under acidic conditions. For example, an acid-labile linker 0630. For example, R is absent. that is hydrolysable in the lysosome or endoSome (e.g., a 0631 For example, R’ is absent. hydrazone, semicarbazone, thiosemicarbazone, cis-aconitic 0632 For example, each of XP and X, independently is amide, orthoester, acetal, ketal, or the like) can be used. Such -O-, - S - N(R') , or absent, in which R' is hydro linkers are relatively stable under neutral pH conditions, such gen, an aliphatic, heteroaliphatic, carbocyclic, or heterocy as those in the blood, but are unstable at below pH 5.5 or 5.0, cloalkyl moiety, C(=O)R', C(=O)CR', SOR' the approximate pH of the lysosome. In certain embodiments, or —N(R')— is a heterocycloalkyl moiety, wherein R' is the hydrolysable linker is a thioether linker (such as, e.g., a hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or het thioether attached to the therapeutic agent via an acylhydra erocycloalkyl moiety. Zone bond. 0633 For example, each of YP, Y, ZP, Z, QP, and Q, 0621. In other embodiments the linker L’or L is photo independently, is absent or a biodegradable linker moiety labile and is useful at the body surface and in many body selected from the group consisting of —S—S—, —C(=O) cavities that are accessible to light. Furthermore, L' or L is biocleavable by infrared light which can penetrate tissue. Accordingly, L' or L is useful for both applications on the body Surface and in the tissue. 0622. In some embodiments, the linker LP or L is bio cleavable by a cleaving agent that is present in the intracellu lar environment (e.g., within a lysosome or endosome or caveolea). The linker can be, for example, a peptidyl linker that is cleaved by an intracellular peptidase or protease enzyme, including, but not limited to, a lysosomal or endo Somal protease. US 2013/0309192 A1 Nov. 21, 2013
NRSO , NRNR C(=O)NRNR , -continued NRNRC(=O) OC(=O)NRNR - NRNRC (—O)O C(—S)NRNR NRNRC(—S) , C(-NR)NRNR , NRNRC(—NR) , O(N–CR) , (CR-N)O C(=O)NR (N=CR) , -(CR—N) NRC(=O) , -SO, , -NRSONR , -SONR , and polyamide, wherein each occurrence of R. R. and Rindependently is hydrogen or an aliphatic, heteroaliphatic, carbocyclic, or heterocyclic moiety, or each occurrence of NR or NRNR is a heterocycloalkyl moiety. 0634) For example, each of MP, MP, MP, MP, M., M', M and M' , independently, is absent or a non-biode gradable linker moiety selected from the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl, heteroalkyl, heteroalk enyl, heteroalkynyl, heterocycloalkyl, aryl, heteroaryl, and a combination thereof and each of MP, MP, MP, M', M°, and M optionally contains one or more —(C=O)—but does not contain any of the biodegradable linker moieties mentioned above. 0635. For example, each of MP, MP, MP, MP, M", M', M and M, independently is Calkyl, Calkyl-C (O) Co. alkyl, Ce alkyl-NH Cole alkyl, Ce alkyl-O- Co. alkyl, C. alkyl-S Co alkyl, C. alkyl-C(O)—C. alkyl-NH, C alkyl-C(O)—C alkyl-O, C-alkyl-C(O)— C. alkyl-S, Co cycloalkyl-C(O)—Cole alkyl, 3-19 mem bered heterocycloalkyl-C(O)—Cole alkyl, aryl-C(O)—Co. alkyl, (CH2CH2O), and the like. 0636. For example, for each L. M' is not absent when O XP is absent. 0637 For example, for each L. M' is not absent when X is absent. Grry 0638 For example, for each LP, at least one of X,Y,ZP, N CH3 s and Q’ is not absent. O 0639 For example, for each L. at least one of X,Y,Z. and Q is not absent. 0640 For example, each of MP' and M' independently is C. alkyl or C. heteroalkyl. 21 pS -- and (0641) For example, each of MPP, MP, MP, M. M., and M, independently is absent, Calkyl, cycloalkyl, het N CH3 O eroalkyl, heterocycloalkyl, or a combination thereof. (0642 For example, for each LP, at most two of MP, MP, and Mare absent. 0643 For example, for each L. at most two of M', M. and Mare absent. 0644. For example, for each LP, one of M' and MP has one of the following structures: in which q is an integer from 0 to 12 and each of p and t independently is an integer from 0 to 3, and the other of M' or MP is either absent or a moiety different from the above, Such as C. alkyl. (0645. For example, for each L, one of M and M has one of the following structures:
O
O p NN su/o, a x-r O US 2013/0309192 A1 Nov. 21, 2013 72
-continued —ZP-MP, or -MP-ZP , independently has one of the fol lowing structures:
(1)
(2)
(3) 2
(4)
(5)
O 4N p ry N CH3 s (6) O
p 21 S ul- and (7) N CH O
(8) in which q is an integer from 0 to 12 and each of p and t independently is an integer from 0 to 3, and the other of M or M is either absent or a moiety different from the above, Such as C. alkyl. (9) 0646 For example, p is 2. (0647 For example, q is 0 or 12. 0648 For example, t is 0 or 1. 0649) For example, each of -MP-ZP , -ZP-MP-, US 2013/0309192 A1 Nov. 21, 2013 73
-continued -continued (10) (19) O > On-O N s H O P-Ph M \, Ph (20) (11) Rld O
(12) O (21) O O O 3. o1 No (22) Rld
O N (13) N1 O O
NY O O (23)
14 O O O (14) Rld /1 Y V
(15) \ 3. (24)
(16)
(17) in which ring A or B independently is cycloalkyl or hetero O cycloalkyl, R' is analiphatic, heteroaliphatic, carbocyclic, or heterocycloalkyl moiety; R' is hydrogen, an aliphatic, het eroaliphatic, carbocyclic, or heterocycloalkyl moiety; and NHN ring D is heterocycloalkyl. 0650 For example, each of -M-Z –Z-M-, —Z-M.--, and -M-Z independently, has one of the O (18) following structures:
(1) RW, >, S SC, s US 2013/0309192 A1 Nov. 21, 2013 74
-continued -continued
(2) f (11)
N N ; and x - S%. .
(3) (12)
x -/ NN2 N (4) >, N
NH in which ring A is cycloalkyl or heterocycloalkyl and R' is hydrogen, an aliphatic, heteroaliphatic, carbocyclic, or het erocycloalkyl moiety. 0651. For example, ring A is 5-19 membered heterocy HO cloalkyl, e.g., (5) C. ^ (6) 0652 For example, ring A is Cls cycloalkyl. i" E. E. E. 5. Ds playin' or piperidinyl. - N 0655 Foror example, R'K. 1Sis U-hydrogen alKyl. or C. alkyl. 0656. For example, ZP is (7)
O
(8)
(9) S 0657 For example, Z is
N
Rld (10) O
H
P-Ph M \, US 2013/0309192 A1 Nov. 21, 2013 75
0658. For example, XP is absent, O or NH. 0661 For example, each of Y and Y independently is 0659 For example, X is absent, O or NH. S—S , COO , COO , CONH O 0660 For example, each of XP and X, independently is NHCO . 0662 For example, each of Q' and Q independently is p absent, S–S COO , -COO – CONH , N Y. OC NHCO. , OCONHNH , or NHNHCOO . O \ / 10663 For example, -LP-D can have one of the following N structures below, in which the wavy bond indicates that D (i.e. Drug) is either connected to the functional linker directly or via another moiety:
(1) (2) O O O O
us R80~. N O Drug s us R80~. N 1 Drug, O O (3) (4) O O O O O us R80 N 1N1N R32ls o Drug, us R1 NH so Drug, O
(5) (6) O O
us N Drug, Drug s
(7) (8) O O O
(9)
(10) (11)
(12) US 2013/0309192 A1 Nov. 21, 2013 76
-continued (13)
O O
NH
HN- O (14) O
O O O H H Drug, R80 N N ~~ NH NH O O O
HN H 1s
(15) O
O O O O H Drug, N R N N 80 H H O
HN HN 1s,
(16) O O O
N R80 N H Drug and O
HN HN 1s, US 2013/0309192 A1 Nov. 21, 2013 77
-continued (17) O O -y H Drug:9. H N R80 N-n N N Y ~~ OT H O H O O
HN HN 1s. wherein Rso is CH2, —NH, or oxygen; and Rs is —NH or oxygen. 0664) For example, polymeric carrier-L-PBRM can have one of the following structures below:
(1)
(2)
O O 1N1 ON1so N PBRM, R80 N 1N1 S 2 O (3) (4)
(5)
(6)
(7) (8)
US 2013/0309192 A1 Nov. 21, 2013 79
-continued O O O (20) --~~-en-)s, --- S PBRM; O ------O O O (21) 22 O O us R30~. -n-ro-n-YNN
wherein: R80 is CH2, NH or oxygen; R8 is
0665 While biocleavable linkers preferably are used in 0670. In certain other embodiments, one or more occur the invention, a non-biocleavable linker also can be used to rences of PBRM is attached to the polymeric carrier, wherein generate the above-described conjugate. A non-biocleavable the one or more occurrences of PBRM may be the same or linker is any chemical moiety that is capable of linking a drug different. In certain other embodiments, one or more polymer or PBRM, to a polymer in a stable, covalent manner. Thus, carriers that contains one or more occurrences of Dare con non-biocleavable linkers are substantially resistant to acid induced cleavage, light-induced cleavage, peptidase-induced nected to a PBRM (e.g., an antibody). cleavage, esterase-induced cleavage, and/or disulfide bond 0671 As discussed more generally above, in certain cleavage, at conditions under which the drug or polymer embodiments, each polymeric carrier independently, has remains active. about 0.1 to about 25% monomers comprising a D. more 0666. In one embodiment, a substantial amount of the drug preferably about 0.5 to about 20%, more preferably about 1 to moiety is not cleaved from the conjugate until the protein about 15%, and even more preferably about 2 to about 10%. polymer-drug conjugate enters a cell with a cell-surface receptor specific for the PBRM of the protein-polymer-drug 0672. In certain embodiments, the conjugate of this inven conjugate, and the drug moiety is cleaved from the protein tion is of Formula (I): polymer-drug conjugate when the protein-polymer-drug con jugate does enter the cell. 0667. In another embodiment, the bioavailability of the protein-polymer-drug conjugate or an intracellular metabo lite of the protein-polymer-drug conjugate in a subject is O O improved when compared to a drug compound or conjugate comprising the drug moiety of the protein-polymer-drug con jugate, or when compared to an analog of the compound not C pi having the drug moiety. O O O l 0668. In another embodiment, the drug moiety is intracel lularly cleaved in a subject from the protein-polymer-drug o={XD conjugate, or an intracellular metabolite of the protein-poly OH OH mer-drug conjugate. MP1Ny D Conjugates or Polymeric Scaffolds MP2 0669 Conjugates of the invention comprise one or more Y wild occurrences of D, where D is a therapeutic agent, e.g., a drug, wherein the one or more occurrences of D may be the same or different. US 2013/0309192 A1 Nov. 21, 2013 80
-continued so that it contains a functional group that can react with a O O functional group of the drug or its derivative; (2) reacting the modified polymer with the drug or its derivative so that the drug is linked to the polymer; (3) modifying the polymer-drug conjugate so that the polymer contains a functional group that CO na can react with a functional group of the PBRM or its deriva tive; and (4) reacting the modified polymer-drug conjugate O ={ with the PBRM or its derivative to form the conjugate of this XD invention. Step (3) may be omitted if the modified polymer MP1 produced by step (1) contains a functional group that can react NyD with a functional group of the PBRM or its derivative. 0678. In another embodiment the conjugates are formed in MP2 several steps: (1) modifying a polymer so that it contains a functional group that can react with a functional group of a MP3N first drug or its derivative; (2) reacting the modified polymer g with the first drug or its derivative so that the first drug is MP4 linked to the polymer; (3) modifying the polymer-drug con ND jugate so that it contains a different functional group that can O O react with a functional group of a second drug or its derivative (4) reacting the modified polymer-drug conjugate with the second drug or its derivative so that the second drug is linked to the polymer-drug conjugate; (5) modifying the polymer CO ria drug conjugate containing 2 different drugs so that the poly mer contains a functional group that can react with a func O ={ tional group of the PBRM or its derivative; and (6) reacting XP the modified polymer-drug conjugate of step (5) with the MPI PBRM or its derivative to form the conjugate of this inven NP tion. Steps (5) and (6) may be repeated if 2 different PBRMor MP2 its derivatives are to be conjugated to form a polymer-drug a. WP conjugate comprising two different drugs and two different O O PBRMS. 0679. In yet another embodiment, the conjugates are formed in several steps. These steps include (1) modifying a polymer so that it contains a functional group that can react O - 14 with a functional group of the drug or its derivative; (2) further modifying the polymer so that it also contains a functional O ={ group that can react with a functional group of the PBRM or XP its derivative; (3) reacting the modified polymer with the drug \ Pl M or its derivative so that the drug is linked to the polymer; and NP (4) reacting the modified polymer-drug conjugate with the V PBRM or its derivative to form the conjugate of this inven MP2 tion. The sequence of steps (1) and (2) or that of steps (3) and Z.l (4) can be reversed. Further either step (1) or (2) may be n MP3 omitted if the modified polymer contains a functional group that can react with both a functional group of the drug or its derivatives and a functional group of the PBRM or its deriva MP4 tive. 0680 In another embodiment the conjugates are formed in PBRM, several steps: (1) modifying a polymer so that it contains a functional group that can react with a functional group of a wherein: first drug or its derivative; (2) further modifying a polymer So each of n, n, n, n, and n is the molar fraction of the that it contains a functional group that can react with a func corresponding polymer unit ranging between 0 and 1: n+n+ tional group of the PBRM or its derivative; (3) reacting the n+n+n-1; provided that none of n, n, and n is 0. modified polymer with the first drug or its derivative so that 0673 For example, the ratio between n and n is greater the first drug is linked to the polymer; (4) modifying the than 1:1 and s200:1. polymer-drug conjugate so that it contains a different func 0674) For example, the ratio between n and n is between tional group that can react with a functional group of a second 10:1 and 50:1. drug or its derivative (5) reacting the modified polymer-drug 0675 For example, the ratio between n and n is between conjugate with the second drug or its derivative so that the 30:1 and 50:1. second drug is linked to the polymer-drug conjugate; (6) 0676 For example, the ratio between n and n is about reacting the modified polymer-drug conjugate containing 2 50:1, 25:1, 10:1, 5:1 or 2:1. different drugs so that the polymer with the PBRM or its 0677. In certain embodiments, the conjugates are formed derivative to form the conjugate of this invention. Step (6) in several steps. These steps include (1) modifying a polymer may be repeated if 2 different PBRM or its derivatives are to US 2013/0309192 A1 Nov. 21, 2013 be conjugated to form a polymer-drug conjugate comprising are predicted to have low toxicity and bioadhesivity, which two different drugs and two different PBRMs. Step (4) may makes them Suitable for several biomedical applications. be carried out after step (1) so that the modified polymer 0683. In certain embodiments of the present invention, the contains two different functional groups that can react with biodegradable biocompatible conjugates can form linear or two different drugs or their derivatives. In this embodiment, branched structures. For example, the biodegradable biocom the modified polymer containing two different functional patible polyal conjugates of the present invention can be group that can react with two different drugs or their deriva chiral (optically active). Optionally, the biodegradable bio tives can be further modified so that it contains a functional compatible polyal conjugates of the present invention can be scalemic. group that can react with a functional group of the PBRM or 0684. In certain embodiments, the conjugates of the inven its derivative; prior to the reaction of the modified polymer tion are water-soluble. In certain embodiments, the conju with either the two different drugs (step (3) and step (5) or gates of the invention are water-insoluble. In certain embodi PBRM (step (6). ments, the inventive conjugate is in a solid form. In certain 0681. The biodegradable biocompatible conjugates of the embodiments, the conjugates of the invention are colloids. In invention can be prepared to meet desired requirements of certain embodiments, the conjugates of the invention are in biodegradability and hydrophilicity. For example, under particle form. In certain embodiments, the conjugates of the physiological conditions, a balance between biodegradability invention are in gel form. and stability can be reached. For instance, it is known that 0685. This invention also features a polymeric scaffold molecules with molecular weights beyond a certain threshold useful for conjugating with a PBRM to form a polymer-drug (generally, above 40-100 kDa, depending on the physical PBRM conjugate described herein. The scaffold comprises a shape of the molecule) are not excreted through kidneys, as polymeric carrier, one or more-LP-D connected to the poly small molecules are, and can be cleared from the body only meric carrier, and one or more L connected to the polymeric through uptake by cells and degradation in intracellular com carrier which is suitable for connecting a PBRM to the poly partments, most notably lysosomes. This observation exem meric carrier, wherein: plifies how functionally stable yet biodegradable materials 0686 each occurrence of D is independently a therapeutic may be designed by modulating their stability under general agent having a molecular weight s5 kDa, physiological conditions (pH=7.5+0.5) and at lysosomal pH 0687 the polymeric carrier is a polyacetal or polyketal, (pH near 5). For example, hydrolysis of acetal and ketal 10688 LP is a linker having the structure: groups is known to be catalyzed by acids, therefore polyals will be in general less stable in acidic lysosomal environment than, for example, in blood plasma. One can design a test to compare polymer degradation profile at, for example, pH-5 and pH-7.5 at 37°C. in aqueous media, and thus to determine the expected balance of polymer stability in normal physi ological environment and in the “digestive’ lysosomal com with R' connected to an oxygen atom of the polymeric partment after uptake by cells. Polymer integrity in Such tests carrier and L' connected to D, and can be measured, for example, by size exclusion HPLC. One skilled on the art can select other suitable methods for study ing various fragments of the degraded conjugates of this invention. 0682. In many cases, it will be preferable that at pH-7.5 the effective size of the polymer will not detectably change over 1 to 7 days, and remain within 50% from the original for at least several weeks. At pH-5, on the other hand, the poly denotes direct or indirect attachment of D to LP', and LP mer should preferably detectably degrade over 1 to 5 days, contains a biodegradable bond so that when the bond is bro and be completely transformed into low molecular weight ken, D is released from the polymeric carrier in an active form fragments within a two-week to several-month time frame. for its intended therapeutic effect; Although faster degradation may be in Some cases preferable, 0689 LP is a carbonyl-containing moiety; in general it may be more desirable that the polymer degrades (0690 L is a linker different from LP and having the struc in cells with the rate that does not exceed the rate of metabo ture: - R' C(=O)-Lif with R' connected to an oxygen lization or excretion of polymer fragments by the cells. atom of the polymeric carrier and L' suitable for connecting Accordingly, in certain embodiments, the conjugates of the directly or indirectly to a PBRM: present invention are expected to be biodegradable, in par (0691 each of R'' and R' independently is absent, alkyl, ticular upon uptake by cells, and relatively “inert” in relation heteroalkyl, cycloalkyl, or heterocycloalkyl, and to biological systems. The products of carrier degradation are (0692 L' is a moiety containing a functional group that is preferably uncharged and do not significantly shift the pH of capable of forming a covalent bond with a functional group of the environment. It is proposed that the abundance of alcohol a PBRM. groups may provide low rate of polymer recognition by cell 0693 For example, L is a linker having the structure: receptors, particularly of phagocytes. The polymer back bones of the present invention generally contain few, if any, antigenic determinants (characteristic, for example, for some polysaccharides and polypeptides) and generally do not com prise rigid structures capable of engaging in "key-and-lock” type interactions in vivo unless the latter are desirable. Thus, the soluble, crosslinked and solid conjugates of this invention US 2013/0309192 A1 Nov. 21, 2013
in which L is a moiety containing a functional group that is wherein: capable of forming a covalent bond with a functional group of 0702 m is an integer from 1 to about 2200, a PBRM, and 0703 m is an integer from 1 to about 660, 0704 m is an integer from 1 to about 300, 0705 m is an integer from 1 to about 110, and 0706 the Sum of m, m, m and m ranges from about 15 to about 2200. (0707 For example, when the PHF in Formula (Ia) has a molecular weight ranging from about 2 kDa to about 40 kDa (i.e., the sum of m, m, m, and m ranging from about 15 to about 300), m is an integer from 1 to about 40, m is an integer from 1 to about 18, and/or m is an integer from 1 to about 140 (e.g., m being about 1-90). (0708 For example, when the PHF in Formula (Ia) has a denotes direct or indirect attachment of L'' to L'. molecular weight ranging from about 6 kDa to about 20 kDa 0694 For example, the functional group of L' or L is (i.e., the sum of m, m, m, and m ranging from about 45 to selected from —SRP. —S-S-LG, maleimido, and halo, in about 150), m is an integer from 2 to about 20, m is an which LG is a leaving group and RP is Hora Sulfur protecting integer from 1 to about 9, and/or m is an integer from 1 to group. about 75 (e.g., m being about 4-45). 0695 For example, L' comprises - X-(CH), C (0709 For example, when the PHF in Formula (Ia) has a (=O)— with X directly connected to the carbonyl group of molecular weight ranging from about 8 kDa to about 15 kDa (i.e., the Sum of m, m, m, and m ranging from about 60 to R-C(=O), in which X is CH, O, or NH, and v is an about 110), m is an integer from 2 to about 15, m is an integer from 1 to 6. integer from 1 to about 7, and/or m is an integer from 1 to (0696. For example, L' or L* contains a biodegradable about 55 (e.g., m being about 4-30). bond. 0710 For example, when the PHF in Formula (Ia) has a (0697 For example, each of R'' and R' is absent. molecular weight ranging from 20 kDa to 300 kDa (i.e., the 0698. For example, the polymeric carrier of the scaffold of Sum of m, m, m, and m ranging from about 150 to about the invention is a polyacetal, e.g., a PHF having a molecular 2200), m is an integer from 3 to about 300, m is an integer weight (i.e., MW of the unmodified PHF) ranging from about from 1 to about 110, and/or m is an integer from 1 to about 2 kDa to about 300 kDa. The selection of a polymeric carrier 660 (e.g., m being about 10-250). with a specific MW range may depend on the size of the 0711 For example, when the PHF in Formula (Ia) has a PBRM to be conjugated with. molecular weight ranging from 40 kDa to 150 kDa (i.e., the 0699 For example, for conjugating a PBRM having a sum of m, m, m, and m ranging from about 300 to about molecular weight of 40 kDa or greater (e.g., 80 kDa or 1100), m is an integer from 4 to about 150, m is an integer from 1 to about 75, and/orm is an integer from 1 to about 330 greater), the polymeric carrier of the scaffold of the invention (e.g., m being about 15-100). is a polyacetal, e.g., a PHF having a molecular weight (i.e., 0712 For example, when the PHF in Formula (Ia) has a MW of the unmodified PHF) ranging from about 2 kDa to molecular weight ranging from about 50 kDa to about 100 about 40 kDa (e.g., about 6-20 kDa or about 8-15 kDa). kDa (i.e., the Sum of m, m, m2, and ms ranging from about 0700 For example, for conjugating a PBRM having a 370 to about 740), m is an integer from 5 to about 100, m is molecular weight of 200 kDa or less (e.g., 80 kDa or less), the an integer from 1 to about 40, and/or m is an integer from 1 polymeric carrier of the scaffold of the invention is a polyac to about 220 (e.g., m being about 15-80). etal, e.g., a PHF having a molecular weight (i.e., MW of the 0713 For example, the scaffold further comprises a unmodified PHF) ranging from about 20 kDa to about 300 PBRM connected to the polymeric carrier via L. kDa (e.g., about 40-150 kDa or about 50-100 kDa). 0714 For example, one or more PBRMs are connected to 0701 For example, the scaffold is of Formula (Ia): one drug-carrying polymeric carrier.
(Ia) O O O O OH OH C CC iii. h O l O O -( inl O ( in2 o i3 LDI --LD LDI D LP2 US 2013/0309192 A1 Nov. 21, 2013 83
0715 For example, the scaffold (e.g., a PBRM-polymer drug conjugate) is of Formula (Ib):
(Ib)
OHr O OHr O O-( i3 o- in4 to for LDI LDI D LP2 LP2
PBRM
wherein: 0723. For example, when the PHF in Formula (Ib) has a molecular weight ranging from 40 kDa to 150 kDa (i.e., the sum of m. m. m. m., and m ranging from about 300 to about 1100), m is an integer from 4 to about 150, m is an integer from 1 to about 75, m is an integer from 1 to about 30, and/or m is an integer from 1 to about 330 (e.g., m being about 10-330 or about 15-100). 0724 For example, when the PHF in Formula (Ib) has a between L- and PBRM denotes directorindirect attachment molecular weight ranging from about 50 kDa to about 100 of PBRM to L', each occurrence of PBRM independently kDa (i.e., the sum of m, m1, m2, m3, and m ranging from has a molecular weight of less than 200 kDa, about 370 to about 740), m is an integer from 5 to about 100, m is an integer from 1 to about 40, m is an integer from 1 to 0716 m is an integer from 1 to about 2200, about 20, and/orm is an integer from 1 to about 220 (e.g. m. 0717 m is an integer from 1 to about 660, being about 15-80). 0718 m is an integer from 3 to about 300, 0725 Alternatively or additionally, one or more drug-car 0719 m is an integer from 0 to about 110, rying polymeric carriers are connected to one PBRM. For 0720 m is an integer from 1 to about 60; and example, the scaffold (e.g., a PBRM-polymer-drug conju 0721 the sum of m. m. m., m and m ranges from about gate) comprises a PBRM with a molecular weight of greater 150 to about 2200. than 40 kDa and one or more D-carrying polymeric carriers 0722. For example, in Formula (Ib), m is an integer from connected to the PBRM, in which each of the D-carrying about 10 to about 660 (e.g., about 10-250). polymeric carrier independently is of Formula (Ic):
(Ic)
OH rY O-( in4 LDI LDI LDI
D LP2 LP2 US 2013/0309192 A1 Nov. 21, 2013
wherein: 074.0 L is a linker different from -R-C(=O)-LP', 0726 terminal and having the structure:-R' C(=O)-Lif with R' con nected to an oxygen atom of the polymeric carrier and L' suitable for connecting to a PBRM; 0741 each of R'' and R' independently is absent, alkyl, heteroalkyl, cycloalkyl, or heterocycloalkyl; 10742 L' is a moiety containing a functional group that is capable of forming a covalent bond with a functional group of D, and attached to L denotes director indirect attachment of L to 10743. L' is a moiety containing a functional group that is PBRM such that the D-carrying polymeric carrier is con capable of forming a covalent bond with a functional group of nected to the PBRM, a PBRM. 0727 m is an integer from 1 to 300, 0744. For example, the D-free scaffold useful to conjugate 0728 m is an integer from 1 to 140, with a PBRM and a D can have one or more of the following 0729 m is an integer from 1 to 40, features. 0730 m is an integer from 0 to 18, 0745) For example, L is a linker having the structure: 0731 m is an integer from 1 to 10; and 0732 the sum of m. m. m. m., and m ranges from 15 to 300; provided that the total number of L° attached to the PBRM is 10 or less. 0733 For example, in Formula (Ic), m is an integer from 1 to about 120 (e.g., about 1-90) and/or m is an integer from 1 to about 10 (e.g., about 1-8). (0734) For example, when the PHF in Formula (Ic) has a in which L is a moiety containing a functional group that is molecular weight ranging from about 6 kDa to about 20 kDa capable of forming a covalent bond with a functional group of (i.e., the Sum of m, m1, m2, m3, and maranging from about 45 a PBRM, and to about 150), m is an integer from 2 to about 20, m is an integer from 1 to about 9, and/or m is an integer from 1 to about 75 (e.g., m being about 4-45). (0735. For example, when the PHF in Formula (Ic) has a molecular weight ranging from about 8 kDa to about 15 kDa (i.e., the Sum of m, m. m. m., and m ranging from about 60 to about 110), m is an integer from 2 to about 15, m is an integer from 1 to about 7, and/or m is an integer from 1 to denotes direct or indirect attachment of L° to L'. about 55 (e.g., m being about 4-30). 10746) For example, the functional group of L' or L is 0736. In another aspect, the invention features a polymeric selected from —SRP. —S S-LG, maleimido, and halo, in scaffold useful to conjugate with both a protein based recog which LG is a leaving group and RP is Hora Sulfur protecting nition-molecule (PBRM) and a therapeutic agent (D). The group. D-free scaffold comprises a polymeric carrier, one or more Lif 0747 For example, L' comprises —X—(CH), C connected to the polymeric carrier which is suitable for con (=O)— with X directly connected to the carbonyl group of necting a PBRM to the polymeric carrier, and one or more R-C(=O), in which X is CH, O, or NH, and v is an —R'' C(=O)-LP connected to the polymeric carrier via integer from 1 to 6. R'', wherein: 0748. For example, L' or L* contains a biodegradable 0737 the polymeric carrier is a polyacetal or polyketal, bond. (0738) R' is connected to an oxygenatom of the polymeric 0749 For example, each of R'' and R' is absent. carrier, (0750 For example, the polymeric carrier of the D-free (0739 L' is a linker suitable for connecting a D molecule scaffold is a polyacetal, e.g., a PHF having a molecular to the polymeric carrier, in which each occurrence of D is weight (i.e., MW of the unmodified PHF) ranging from about independently a therapeutic agent having a molecular weight 2 kDa to about 300 kDa. s5 kDa; (0751. The D-free scaffold is of Formula (Id):
to notLDI toL otho LP2 US 2013/0309192 A1 Nov. 21, 2013
wherein: wherein: 0752 m is an integer from 1 to about 2200, 0753 m is an integer from 1 to about 660, 0754 m is an integer from 1 to about 110, and 0755 the sum of m, m, and m ranges from about 15 to about 2200. (0756. For example, when the PHF in Formula (Id) has a molecular weight ranging from about 2 kDa to about 40 kDa (i.e., the sum of m, m, and m ranging from about 15 to about between L- and PBRM denotes director indirect attachment 300), m is an integer from 1 to about 18, and/or m is an of PBRM to L, PBRM has a molecular weight of less than integer from 1 to about 140 (e.g., m being about 2-120). 200 kDa, (0757. For example, when the PHF in Formula (Id) has a molecular weight ranging from about 6 kDa to about 20 kDa 0765 m is an integer from 1 to 2200, (i.e., the sum of m, m, and m ranging from about 45 to about 0766 m is an integer from 1 to 660, 150), m is an integer from 1 to about 9, and/orm is an integer from 1 to about 75 (e.g., m being about 6-60). 0767 m is an integer from 0 to 110, (0758. For example, when the PHF in Formula (Id) has a 0768 m is an integer from 1 to about 60; and molecular weight ranging from about 8 kDa to about 15 kDa 0769 the sum of m, m, m, m- and m ranges from about (i.e., the sum of m, m, and m ranging from about 60 to about 150 to about 2200. 110), m is an integer from 1 to about 7, and/orm is an integer from 1 to about 55 (e.g., m being about 6-45). 0770 For example, in Formula (Ie), m is an integer from (0759 For example, when the PHF in Formula (Id) has a about 10 to about 660 (e.g., about 14-550). molecular weight ranging from 20 kDa to 300 kDa (i.e., the (0771) For example, when the PHF in Formula (Ie) has a Sum of m, m, and m ranging from about 150 to about 2200), molecular weight ranging from 40 kDa to 150 kDa (i.e., the m is an integer from 1 to about 110, and/or m is an integer Sum of m, m, m, and m ranging from about 300 to about from 1 to about 660 (e.g., m being about 13-550). 1100), m is an integer from 1 to about 75, m is an integer 0760 For example, when the PHF in Formula (Id) has a from 1 to about 30, and/orm is an integer from 1 to about 330 molecular weight ranging from 40 kDa to 150 kDa (i.e., the (e.g., m being about 20-250). Sum of m, m, and m ranging from about 300 to about 1100), m is an integer from 1 to about 75, and/or m is an integer (0772 For example, when the PHF in Formula (Ie) has a from 1 to about 330 (e.g., m being about 20-250). molecular weight ranging from about 50 kDa to about 100 0761 For example, when the PHF in Formula (Id) has a kDa (i.e., the Sum of m, m, m3, and m ranging from about molecular weight ranging from about 50 kDa to about 100 370 to about 740), m is an integer from 1 to about 40, m is kDa (i.e., the Sum of m, m, and m ranging from about 370 to an integer from 1 to about 20, and/or m is an integer from 1 about 740), m is an integer from 1 to about 40, and/or m is to about 220 (e.g., m being about 20-180). an integer from 1 to about 220 (e.g., m being about 20-180). 0773 Alternatively or additionally, one or more D-free 0762. For example, the D-free scaffold further comprises a polymeric carriers are connected to one PBRM. For example, PBRM connected to the polymeric carrier via L. the scaffold comprises a PBRM with a molecular weight of 0763 For example, one or more PBRMs are connected to greater than 40 kDa and one or more polymeric carriers one D-free polymeric carrier. connected to the PBRM, in which each of the polymeric 0764 For example, the D-free scaffold is of Formula (Ie): carrier independently is of Formula (Ih):
to theLDI othoLD US 2013/0309192 A1 Nov. 21, 2013 86
(Ih) O O O O
OH O O O ( i3 O in4 LDI LDI to to LDI
LP2
wherein: (i.e., the sum of m, m, m, and m ranging from about 45 to 0774 terminal about 150), m is an integer from 1 to about 9, and/orm is an integer from 6 to about 75 (e.g., m being about 7-60). 0782. For example, when the PHF in Formula (Ih) has a molecular weight ranging from about 8 kDa to about 15 kDa (i.e., the Sum of m, m, m, and m ranging from about 60 to about 110), m is an integer from 1 to about 7, and/orm is an integer from 6 to about 55 (e.g., m being about 7-45). attached to L denotes director indirect attachment of L° to 0783 PBRM-drug-polymer conjugates, drug carring PBRM such that the D-carrying polymeric carrier is con polymeric scaffolds, or PBRM-caning polymer scaffolds can nected to the PBRM, be purified (i.e., removal of residual unreacted drug, PBRM, (0775 m is an integer from 1 to 300, or polymeric starting materials) by extensive diafiltration. If 0776 m is an integer from 1 to 140, necessary, additional purification by size exclusion chroma (0777 m is an integer from 0 to 18, tography can be conducted to remove any aggregated PBRM 0778 m is an integer from 1 to 10; and drug polymer conjugates. In general, the PBRM-drug poly 0779 the sum of m, m, m, and maranges from 15 to 300; mer conjugates as purified typically contain (5% aggregated provided that the total number of L'attached to the PBRM is PBRM-drug polymer conjugates as determined by SEC or 10 or less SDS-PAGE: <1% polymer-drug conjugate as determined by 0780. For example, in Formula (Ih), m is an integer from SEC and <2% unconjugated PBRM as determined by RP 2 to about 130 (e.g., about 3-120) and/or m is an integer from HPLC. 1 to about 10 (e.g., about 1-8). 0784 Tables D and Ebelow provide examples of the drug 0781. For example, when the PHF in Formula (Ih) has a carrying polymeric scaffolds and the polymer-drug-protein molecular weight ranging from about 6 kDa to about 20 kDa conjugates of the invention respectively. US 2013/0309192 A1 Nov. 21, 2013 87
O
() CIHT8IVIL OH
6XEI US 2013/0309192 A1 Nov. 21, 2013 88
O No.
6XEI US 2013/0309192 A1 Nov. 21, 2013 89
HS
?¡¿ US 2013/0309192 A1 Nov. 21, 2013 90
HO
OHO
iº^~:•~uli DO:DO, (DO,DO,DO HS O H
HO D) () ()O O (DO, US 2013/0309192 A1 Nov. 21, 2013 91
O
HS 3() ? DO:DO, ONIHOH
8]XII