Granted Patent B2

US 9,272,048 · App. 12/990,101 · Granted Mar 1, 2016

Inventors: Harald Rau (Heidelberg, DE); Susanne Kinderman (Heidelberg, DE); Torben Lebmann (Mannheim, DE); Grethe Norskov Rasmussen (Farum, DK); Ulrich Hersel (Heidelberg, DE); Thomas Wegge (Heidelberg, DE); Kennett Sprogoe (Palo Alto, CA)

Assignee: Ascendis Pharma Growth Disorders Division A/S

A61K47/48215A61K47/48338

Abstract

Claims (204)

1. A pharmaceutical composition comprising:

suitable pharmaceutical excipients; and

a human in vivo clinical effective amount of a recombinant human growth hormone (rhGH) PEGylated prodrug conjugate, wherein PEG is linked to rhGH via a self hydrolysable (autocleavage) transient linker from which the rhGH is released in its native form;

wherein the prodrug conjugate has a GH activity which is less than 5% of the native growth hormone without PEG; and

wherein the linker autohydrolysis rate of the prodrug conjugate is such that the in vivo half-life of the prodrug conjugate is from 10 hours to 600 hours;

wherein the term in vivo half-life refers to the time interval in which 50% of the initial proportion of the growth hormone is released from the hGH PEGylated prodrug.

2. The pharmaceutical composition of claim 1 ;

wherein the autohydrolysis rate is such that the in vivo half life is up to 5 times shorter than the corresponding hGH PEGylated prodrug conjugate's in vitro half-life.

3. The pharmaceutical composition of claim 2 ;

wherein the in vivo half life is up to 3 times shorter than the corresponding hGH PEGylated prodrug conjugate's in vitro half-life.

4. The pharmaceutical composition of claim 2 ;

wherein the in vivo half-life is up to 2 times shorter than or almost identical to the corresponding hGH PEGylated prodrug conjugate's in vitro half-life.

5. The pharmaceutical composition of claim 1 ;

wherein the composition is a composition for subcutaneous administration, intramuscular administration or intravenous injection.

6. The pharmaceutical composition of claim 1 ;

wherein the PEGylated prodrug has a total PEG load per growth hormone molecule that amounts to at least 25 kDa.

7. The pharmaceutical composition of claim 1 ;

wherein the prodrug conjugate has the chemical structure (A):

wherein:

HN-rhGH represents the rhGH residue attached to the transient linker;

R1, R2, R3, R4, and R5 are selected independently from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tertiary butyl;

PEG represents the pegylation residue attached to the transient linker;

n=1 or 2; and

X is selected from C1 to C8 alkyl or C1 to C12 heteroalkyl.

8. The pharmaceutical composition of claim 7 ;

wherein the partial structure of formula (I) within structure (A) of claim 7 , which is

is selected from the group consisting of:

9. The pharmaceutical composition of claim 7 ;

wherein the partial structure of formula (II) within structure (A) of claim 7 , which is

is selected from the group consisting of:

10. The pharmaceutical composition of claim 7 ;

wherein PEG-X in formula (A) of claim 7 is PEG-W of formula III:

wherein PEG-W is selected from the group consisting of:

11. The pharmaceutical composition of claim 1 ;

wherein the prodrug conjugate structure is:

12. The pharmaceutical composition of claim 1 ;

wherein the prodrug conjugate structure is:

wherein:

HN-rhGH represents the rhGH residue attached to the transient linker; PEG represents the pegylation residue attached to the transient linker;

R is selected from hydrogen, methyl, ethyl, propyl and butyl; and

X is selected from C1 to C8 alkyl or C1 to C12 heteroalkyl.

13. The pharmaceutical composition of claim 1 ;

wherein the linker autocleavage rate in vivo is such that the in vivo half-life is from 20 to 300 hours.

14. The pharmaceutical composition of claim 1 ;

wherein the linker autocleavage rate in vivo is such that the in vivo half-life is from 20 to 150 hours.

15. The pharmaceutical composition of claim 1 ;

wherein the linker autocleavage rate in vivo is such that the in vivo half-life is from 30 to 100 hours.

16. The pharmaceutical composition of claim 1 ;

wherein the linker autocleavage rate in vivo is such that the in vivo half-life is from 30 to 75 hours.

17. A method for treating a GH related disease in a human person, comprising:

utilizing a clinical effective amount of the pharmaceutical composition of claim 1 .

18. A pharmaceutical composition comprising:

suitable pharmaceutical excipients; and

a prodrug conjugate of the human growth hormone (hGH) of formula (AA):

hGH-NH-L a -S 0   (AA);

wherein:

hGH-NH represents the hGH residue;

L a represents a functional group, which is self hydrolysable (auto-cleavable) by an auto-cleavage inducing group G a ;

S 0 is a polymer chain having a molecular weight of at least 5 kDa and comprising an at least first branching structure BS 1 ; and

the at least first branching structure BS 1 comprises an at least second polymer chain S 1 having a molecular weight of at least 4 kDa;

wherein at least one of S 0 , BS 1 , S 1 further comprises the auto-cleavage inducing group G a ; and

wherein:

the branching structure BS 1 further comprises an at least third polymer chain S 2 having a molecular weight of at least 4 kDa; or

at least one of S 0 , S 1 comprises an at least second branching structure BS 2 comprising the at least third polymer chain S 2 having a molecular weight of at least 4 kDa;

wherein the molecular weight of the prodrug conjugate without the hGH-NH is at least 25 kDa and at most 1000 kDa.

19. The composition of claim 18 ;

wherein the molecular weight of the prodrug conjugate without the hGH-NH is at least 30 kDa and at most 120 kDa.

20. The composition of claim 18 ;

wherein the molecular weight of the prodrug conjugate without the hGH-NH is at least 40 kDa and at most 100 kDa.

21. The composition of claim 18 ;

wherein the molecular weight of the prodrug conjugate without the hGH-NH is at least 40 kDa and at most 90 kDa.

22. The composition of claim 18 ;

wherein L a is selected from the group consisting of C(O)—O— and C(O)—, which together with the primary amino group hGH, form a carbamate or amide group resulting in formula (AA1) or (AA2):

hGH-NH—C(O)O—S 0   (AA1);

hGH-NH—C(O)—S 0   (AA2).

23. The composition of claim 18 ;

wherein L a , together with the amino group of hGH, forms a carbamate functional group, the cleavage of said group being induced by a hydroxyl or amino group of G a via 1,4- or 1,6 benzyl elimination of S 0 ; and

wherein G a contains ester, carbonate, carbamate, or amide bonds that undergo rate-limiting transformation.

24. The composition of claim 18 ;

wherein G a is an aromatic ring or fluorenylmethyl directly attached to a carbamate functional group formed by L a and the primary amino group of hGH.

25. The composition of claim 18 ;

wherein at least one of the branching structures BS 1 , BS 2 comprises a further fourth polymer chain S 3 having a molecular weight of at least 4 kDa; or

one of S 0 , S 1 , S 2 comprises a third branching structure BS 3 comprising the at least fourth polymer chain S 3 having a molecular weight of at least 4 kDa.

26. The composition of claim 18 ;

wherein the at least three chains S 0 , S 1 , S 2 are independently based on a polymer selected from the group consisting of:

polyalkyloxy polymers, hyaluronic acid and derivatives thereof, polyvinyl alcohols, polyoxazolines, polyanhydrides, poly(orthoesters), polycarbonates, polyurethanes, polyacrylic acids, polyacrylamides, polyacrylates, polymethacrylates, polyorganophosphazenes, polysiloxanes, polyvinylpyrrolidone, polycyanoacrylates, and polyesters.

27. The composition of claim 26 ;

wherein the at least three chains S 0 , S 1 , S 2 are based on a polyalkoxy polymer.

28. The composition of claim 18 ;

wherein the shortest distance between the attachment site of S 0 to L a and the first branching structure BS 1 measured as connected atoms is less than 50 atoms.

29. The composition of claim 28 ;

wherein the shortest distance is less than 20 atoms.

30. The composition of claim 18 ;

wherein S 0 is of formula (AAA1):

wherein:

G a has the meaning as indicated in claim 18 ;

S 00 is CH2 or C(O);

S 0A is an alkylene chain having from 1 to 20 carbon atoms, which is optionally interrupted or terminated by one or more groups, cycles, or heteroatoms selected from the group consisting of:

optionally substituted heterocycle, O, S, C(O), and NH;

BS 1 , BS 2 , and BS 3 are independently selected from the group consisting of N and CH;

S 0B and S 1A are independently an alkylene chain having from 1 to 200 carbon atoms, which is optionally interrupted or terminated by one or more groups, cycles, or heteroatoms selected from the group consisting of:

optionally substituted heterocycle, O, S, C(O), and NH;

S 0C and S 1B are (C(O)) n2 (CH 2 ) n1 (OCH 2 CH 2 ) n OCH 3 , wherein:

each n is independently an integer from 100 to 500;

each n1 is independently 0, 1, 2, 3, 4, 5, 6, 7, or 8; and

n2 is 0 or 1;

S 2 and S 3 are independently:

hydrogen; or

(C(O)) n2 (CH 2 ) n1 (OCH 2 CH 2 ) n OCH 3 , wherein:

 each n is independently an integer from 100 to 500;

 each n1 is independently 0, 1, 2, 3, 4, 5, 6, 7, or 8; and

 n2 is 0 or 1;

provided that at least one of S 2 , S 3 is other than hydrogen; and

R 2 and R 3 are selected independently from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tertiary butyl.

31. The composition of claim 30 ;

wherein:

G a is OC(O)—R; and

R is the partial structure of formula (I):

wherein:

R1, R4, and R5 are selected independently from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tertiary butyl; and

n=1 or 2.

32. The composition of claim 18 ;

wherein L a -S 0 is represented by formula (AAA2):

wherein:

the dashed line indicates the attachment to the primary amino group of hGH so that L a and the amino group form an amide bond;

X is C(R 4 R 4a ), N(R 4 ), O, C(R 4 R 4a )—C(R 5 R 5a ), C(R 5 R 5a )—C(R 4 R 4a ), C(R 4 R 4a )—N(R 6 ), N(R 6 )—C(R 4 R 4a ), C(R 4 R 4a )—O, or O—C(R 4 R 4a );

X 1 is C; or S(O);

X 2 is C(R 7 , R 7a ); Or C(R 7 , R 7a )—C(R 8 , R 8a );

R 1 , R 1a , R 2 , R 2a , R 3 , R 3a , R 4 , R 4a , R 5 , R 5a , R 6 , R 7 , R 7a , R 8 , R 8a are independently selected from the group consisting of H and C 1-4 alkyl, with the provisos that:

optionally, one or more of the pairs R 1a /R 4a , R 1a /R 5a , R 4a /R 5a , R 4a /R 5a , R 7a /R 8a form a chemical bond;

optionally, one or more of the pairs R 1 /R 1a , R 2 /R 2a , R 4 /R 4a , R 5 /R 5a , R 7a /R 7a , R 8 /R 8a are joined together with the atom to which they are attached to form a C 3-7 cycloalkyl; or 4 to 7 membered heterocyclyl;

optionally, one or more of the pairs R 1 /R 4 , R 1 R 5 , R 1 R 6 , R 4 /R 5 , R 7 /R 8 , R 2 /R 3 are joined together with the atoms to which they are attached to form a ring A; and

optionally, R 3 /R 3a are joined together with the nitrogen atom to which they are attached to form a 4 to 7 membered heterocycle;

A is selected from the group consisting of phenyl, naphthyl, indenyl, indanyl, tetralinyl, C 3-10 cycloalkyl, 4 to 7 membered heterocyclyl, and 9 to 11 membered heterobicyclyl; and

wherein S 0 is substituted with one group L 2 -Z and optionally further substituted, provided that the hydrogen marked with the asterisk in formula (I) is not replaced by a substituent, wherein:

L 2 is a single chemical bond or a spacer; and

Z is of formula (AAA2a):

wherein:

S 00 is CH2 or C(O);

S 0A is an alkylene chain having from 1 to 20 carbon atoms, which is optionally interrupted or terminated by one or more groups, cycles, or heteroatoms selected from the group consisting of:

optionally substituted heterocycle, O, S, C(O), and NH;

BS 1 , BS 2 , and BS 3 are independently selected from the group consisting of N and CH;

S 0B and S 1A are independently an alkylene chain having from 1 to 200 carbon atoms, which is optionally interrupted or terminated by one or more groups, cycles, or heteroatoms selected from the group consisting of:

optionally substituted heterocycle, O, S, C(O), and NH;

S 0C and S 1B are (C(O)) n2 (CH 2 ) n1 (OCH 2 CH 2 ) n OCH 3 , wherein:

each n is independently an integer from 100 to 500;

each n1 is independently 0, 1, 2, 3, 4, 5, 6, 7, or 8; and

n2 is 0 or 1; and

S 2 and S 3 are independently:

hydrogen; or

(C(O)) n2 (CH 2 ) n1 (OCH 2 CH 2 ) n OCH 3 , wherein:

 each n is independently an integer from 100 to 500;

 each n1 is independently 0, 1, 2, 3, 4, 5, 6, 7, or 8; and

 n2 is 0 or 1;

provided that at least one of S 2 , S 3 is other than hydrogen.

33. A pharmaceutical composition comprising suitable pharmaceutical excipients: and

a prodrug conjugate of the human growth hormone (hGH) of formula (AB):

hGH-(NH-L-S 0 )  (AB);

wherein:

n is 2, 3, or 4;

hGH(—NH)n represents the hGH residue;

each L is independently a permanent functional group L p ; or a functional group L a , which is self hydrolysable (autocleavable) by an autocleavage inducing group G a ; and

each S 0 is independently a polymer chain having a molecular weight of at least 5 kDa;

wherein S 0 is optionally branched by comprising an at least first branching structure BS 1 , the at least first branching structure BS 1 comprising an at least second polymer chain S 1 having a molecular weight of at least 4 kDa; and

wherein at least one of S 0 , BS 1 , S 1 further comprises the auto-cleavage inducing group G a ;

wherein the molecular weight of the prodrug conjugate without the hGH-NH is at least 25 kDa and at most 1000 kDa.

34. The composition of claim 33 ;

wherein n is 2.

35. A prodrug comprising:

a prodrug conjugate of the human growth hormone (hGH) of formula (AA):

hGH-NH-L a -S 0   (AA);

wherein:

hGH-NH represents the hGH residue;

L a represents a functional group, which is self hydrolysable (auto-cleavable) by an auto-cleavage inducing group G a ;

S 0 is a polymer chain having a molecular weight of at least 5 kDa and comprising an at least first branching structure BS 1 ; and

the at least first branching structure BS 1 comprises an at least second polymer chain S 1 having a molecular weight of at least 4 kDa,

wherein at least one of S 0 , BS 1 , S 1 further comprises the auto-cleavage inducing group G a ; and

wherein:

the branching structure BS 1 further comprises an at least third polymer chain S 2 having a molecular weight of at least 4 kDa or

at least one of S 0 , S 1 comprises an at least second branching structure BS 2 comprising the at least third polymer chain S 2 having a molecular weight of at least 4 kDa;

wherein the molecular weight of the prodrug conjugate without the hGH-NH is at least 25 kDa and at most 1000 kDa.

36. A prodrug selected from the group consisting of:

wherein m is an integer from 200 to 250 and n is an integer from 100 to 125;

wherein n is an integer from 400 to 500;

wherein n is an integer from 400 to 500; and

wherein n is an integer from 400 to 500.

37. A method for preparing a compound of formula:

hGH-NH—C(O)O—S 0   (AA1);

wherein:

S 0 is a polymer chain having a molecular weight of at least 5 kDa, and comprising an at least first branching structure BS 1 ; and

the at least first branching structure BS 1 comprises an at least second polymer chain S 1 having a molecular weight of at least 4 kDa;

wherein at least one of S 0 , BS 1 , S 1 further comprises the auto-cleavage inducing group G a ; and

wherein:

the branching structure BS 1 further comprises an at least third polymer chain S 2 having a molecular weight of at least 4 kDa; or

at least one of S 0 , S 1 comprises an at least second branching structure BS 2 comprising the at least third polymer chain S 2 having a molecular weight of at least 4 kDa; and

wherein S 0 comprises at least one group:

the method comprises the following steps:

(a) reacting a compound of formula ROC(O)O—S 0′ —SH (AA1) with a compound of formula (AA2), resulting in a compound of formula ROC(O)O—S 0 , wherein:

formula (AA2) is:

R is pentafluorophenyl or NHS; and

S 0′ and S 0″ are selected to yield S 0 comprising the at least one group:

and

(b) reacting the compound of formula ROC(O)O—S 0 with hGH-NH 2 , to yield a compound of formula (AA1), wherein:

hGH-NH 2 represents hGH with one of its primary amino groups.

Assignments (9)

Priority Claims (3)

Continuity (1)