WO1990007519A1

WO1990007519A1 - 3'-deamino analogs of esorubicin and methods for their use

Info

Publication number: WO1990007519A1
Application number: PCT/US1989/004717
Authority: WO
Inventors: Waldemar Priebe; Nouri Neamati-M; Roman Perez-Soler
Original assignee: University of Texas System; University of Texas at Austin
Current assignee: University of Texas System; University of Texas at Austin
Priority date: 1988-12-28
Filing date: 1989-10-23
Publication date: 1990-07-12
Anticipated expiration: 1991-06-28
Also published as: AU4752190A

Abstract

Compounds having formula (I) have been found to have antitumor activity. R1 is selected from the group consisting of H, OCH¿3?, OH, and F; R?2¿ is selected from the group consisting of H, OH, and carboxy having the formula COOR6; R3 is selected from the group consisting of COCH¿2R?7 and CH¿2?CH2R?7; R4¿ is selected from the group consisting of H, aliphatic and aromatic hydrocarbons having 1 to 10 carbons, and acyl groups having the formula COR8; R5 is selected from the group consisting of CH¿3? and CH2OH; R?6¿ is selected from the group consisting of H and aliphatic hydrocarbons having 1 to 6 carbons; R7 is selected from the group consisting of H, OH, and OR9; R8 is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons; R9 is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons and COR?10; and R10¿ is selected from the group consisting of aliphatic hydrocarbons having 1 to 18 carbons.

Description

3'-DEAMINO ANALOGS OF ESORUBICIN AND METHODS FOR THEIR USE

The U.S. government may own certain rights in this patent pursuant to National Institutes of Health Grant No, RR 5511-23.

The present invention relates to anthracycline antibiotics and methods of using such compounds and formulations thereof in antitumor therapy.

Anthracycline antibiotics such as doxorubicin, esorubicin, and daunorubicin are known to have antineoplastic activity. However, these compounds and many of their derivatives have serious side effects/ such as cardiotoxicity, which severely restrict the dosage and the frequency with which they can be administered, and thus limit their overall effectiveness. There is a long standing need for analogs which will have higher activity and lower toxicity, as well as a broader spectrum of antitumor activity, than the previously known compounds. The present invention includes compounds which have the formula:

R is selected from the group consisting of H, 0CH ,

OH, and F. R 2 is selected from the group consisting of H,

OH, and carboxy having the formula COOR . R is selected

4 . from the group consisting of COCH-R and CH₂CH-R . R is selected from the group consisting of H, aliphatic and aromatic hydrocarbons having from 1 to 10 carbons, and acyl groups having the formula COR 8. R5 is selected from c the group consisting of CH- and CH₂OH. R is selected from the group consisting of H and aliphatic hydrocarbons having from 1 to 6 carbons. is selected from the group c - consisting of H, OH, and OR" R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 g carbons. R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons and COR

R is selected from the group consisting of aliphatic hydrocarbons having 1-18 carbons, preferably 1-6 carbons.

R 1 is preferably 0CH , R2 is preferably H, R3 is preferably COCH-OH or COCH₃, R 4 preferably is H, and R5 is preferably CH_.

"Aliphatic and aromatic hydrocarbons" is used in this patent to mean both unsubstituted and substituted hydro¬ carbons _r such as_r for example, those substituted with acyl or hydroxyl groups. The present invention also concerns compositions which include an effective amount of a compound in accordance with the above formula and a pharmaceutically acceptable carrier.

These compounds and compositions have been found to have antitumor activity, and are useful in methods of inhibiting neoplastic cell growth in a mammal. In a particular embodiment, the present invention concerns methods of inhibiting lymphoid leukemia cell growth. Such methods involve administering to a mammal an effective amount of a compound in accordance with the present invention. Particular compounds in accordance with the present invention have been shown to have high activity.

Figures 1, 2, and 3 show synthetic schemes for preparation of compounds in accordance with the present invention.

Specific anthracycline antibiotics in accordance with the present invention include the following two examples:

Synthesis of such compounds can suitably be done as follows.

l,5-Anhydro-2,6-dideoxy-L-arabino-hex-l-enitol (compound 2, L-rhamnal)

To a solution of 3,4-di-O-acetyl-L-rhamnal (compound 1; Pfanstiehl Labs, Inc.) (57 g, 0.266 mole) in absolute methanol (500 ml) was added with stirring anion-exchange resin (8.5 g, Amberlite IRN-78, OH-form) . The solution was monitored with TLC (hexane-ethyl acetate 2:1) until completion of the reaction (18 hours). The mixture was filtered and then evaporated under diminished pressure. The product was compound 2 (Rf 0.09, hexane-ethyl acetate 2:1), which was crystalized upon evaporation of solvent to give 34.5 g (100%).

3-O-tert-Butyldimethylsilyl-L-rhamnal (compound 3a)

To a solution of compound 2 (34.5 g, 0.265 mol) and imidazole (45 g, 0.663 mol) in N,N-dimethylformamide (150 ml) was added with stirring t-butylchlorodimethyl silane (43.9 g, 0.292 mol). The solution was stirred at room temperature until complete disappearance of the substrate (2 hours). The mixture was then poured into water (300 ml) and extracted with hexane (800 ml x 3). The organic extract was combined and washed with water (500 ml), dried over sodium sulfate and evaporated. The resulting oil (63 g) was used in the next step as a mixture containing additionally compounds 3b (3,4-di-O-tert-butyldimethyl- silyl-L-rhamnal) (1.5%) and 3c (4-O-tert- butyldimethylsilyl-L-rhamnal) (3%). 3-0-tert-Butyldimethylsilyl-4-0-(methylthio)thiocarbonyl- -rhamnal (compound 4)

To a solution in dry oxalane (600 ml) containing compound 3a (63 g, 0.<_58 mol), and a minute amount of commpounds 3b and 3c, and imidazole (63 mg) under argon, was added sodium hydride (15.5 g, 0.387 mol, 60% dispersion in mineral oil). After stirring for 2 hours at room temperature, carbon disulfide (58.9 g, 0.774 mol) was added and stirring continued for two more hours after which iodomethane (64.8 g, 0.456 mol) was added and stirring was continued overnight. Then glacial acetic acid (13 ml) was added slowly to destroy the excess of sodium hydride. Solvent was removed and ether (400 ml) was added and the mixture was washed with 5% NaHCO- (150 ml) and water (150 ml x 2). The organic extract was dried over sodium sulfate, filtered and evaporated. The resulting oil (84 g), was used in the next st^p as a mixture of a major product Rf 0.8 and a minor product Rf 0.88 (hexane-toluene 4:3). ¹³C-NMR 6:215.5(C=S) ,

143.5(C-1), 102.8(C-2), 82.8(C-4), 72.4(C-5), 66.1(C-3), 25.6(Me_₃CSi), 19.2(CH₃S), 17.9(CH₃CSi) , 16.6(C-6).

3-0-tert-Butyldimethylsilyl-4-deoxy-L-rhamnal (compound 5)

To a solution of compound 4 (84 g, 0.251 mol) in anhydrous toluene (500 ml) under argon was added AIBN (2,2*-azobisisobutyronitrile) (4.2 g) and tributyltin hydride (87.7 g, 0.301 mol). The solution was stirred at 90°C until complete disappearance of the substrate (45 min.). The solvent was then removed and the product was chromatographed with hexane-toluene (10:1). Compound 5

13 was obtained as an oil (48.8 g, 85.0% yield). C-NMR

6:144.3(C-1), 105.9(02), 71.0(05), 63.6(03), 39.9(C-4), 25.8(Me₃CSi) , 20.9(06), 18.l(Me,CSi) , -4.62(Me₂Si) . Figure 1 also shows an alternate synthetic scheme for preparing compound 5, starting with 3,4-di-O-acetyl- - fucal instead of 3,4-di-O-acetyl-L-rhamnal.

3-Q-Acetyl-4-deoxy-L-rhamnal (compound 6)

Compound 5 (34.5 g, 0.151 mol) was dissolved in oxalane (300 ml), dichloromethane (150 ml), pyridine (10 ml), and tetrabutylammonium flouride (0.23 mol, 223 ml of a 1M solution in oxalane) was added. The solution was stirred at room temperature until complete disappearance of the substrate (48 hours). Sodium sulfate (anhydrous) was added and the mixture was stirred for an additional 20 minutes and then filtered and evaporated. Pyridine (100 ml) was added followed by acetic anhydride (21.5 g, 0.21 mol) and stirring was continued at room temperature for 4 more hours. Dichloromethane (300 ml) was then added and the mixture was washed with 0.IN HC1 (100 ml), 5% NaHC0₃ (100 ml), and water (3 x 100 ml). The organic layer was dried over sodium sulfate, filtered and evaporated. Compound 6 was obtained as an o l (21.15 g, 89.8%). 13C-

NMR δ: 170.7(C=0), 146.6(01), 100.7(02), 70.7(05),

65.6(03), 35.1(04), 21.0(OAc)_f 20.5{O6). .

7-0-(3-0- cetyl-2,4,6-trideoxγ-α-L-threo-hexopyranosyl) daunomycinone (compound 9)

A solution of 3-0-acetyl-l,5-anhydro-2,4,6-trideoxy- L-threo-hex-1-enitol (compound 6, 235 mg, 1.51 mmol) in dry benzene was treated with dry hydrogen chloride gas for 5 minutes until a more polar product (compound 7; TLC:hexane-ethyl acetate 4:1; Rf 0.1) was formed. Benzene was removed and dichloromethane (10 ml) was added and^' the solution was poured into a suspension of daunomycinone (compound 8, 300 mg, 0.75 mmol), 4A molecular sieves (3.0 g), mercuric bromide (45 mg), and yellow mercuric oxide

BSTITUTESHEET (600 mg) in dichloromethane (50 ml). The resulting mixture was stirred for 60 minutes, after which an addi¬ tional 1.0 equivalent of glycosyl chloride (compound 7, 1.51 mmol) was added, and stirring continued for two more hours until disappearance of substrate. Salts were filtered off and the filtrate was diluted with dichloro¬ methane (100 ml) and extracted with 10% aqueous potassium iodide (50 ml x 2) and water (50 ml x 2). The organic extract was dried over sodium sulfate and evaporated. The obtained mixture was immediately chromatographed on silica gel (toluene-acetone, 20:1) to give after crystallization (dichloromethane-hexane) compound 9 (225 mg, 54%); mp 150-155°C; IRmax(KBr) 3487 (OH), 1737 and 1718(C0), 1617 and 1578 (H-bonded quinone), 1411, 1370, 1285, 1236, 1206, 1121, 1032, and 979 cm^-1.

Analysis: Calculated for ^c ₂o^H ₃₀°_1:L ' ^{L 2} H 0(563.54): C 61.81, H 5.54 Found: C 61.46, H 5.49

7-0-(2,4,6-Trideoxy-α-L-threo-hexopyranosyl)daunomycinone (compound 10)

A solution of compound 9 (120 mg, 0.22 mmol) in methanol (10 ml) was treated with a 0.5 M solution of sodium methoxide in methanol (0.5 ml). After 90 minutes of stirring at room temperature the reaction was completed. Dry ice was added and the mixture was diluted with dichloromethane (100 ml) and washed with water (40 ml x 3), then dried over sodium sulfate, filtered and evaporated. TLC showed one spot having Rf 0.11 (toluene- acetone, 8:1). The product was crystallized from dichloromethane and hexane; yield 83 mg (75%); mp 143- 148^βC; IRmax(KBr) 3470(OH), 1713(CO), 1616 and 1579 (H- bonded quinone), 1411, 1350, 1285, 1207, 1123, 1087, 1031, 991 and 976 cm^-1. Analysis: Calculated for C₂₇H₂₈O₁₀ ^* 1/2 H₂(521.5):

C 62.18 H 5.60 Found: C 61.79 H 5.40

14-0-tert-Butyldiιnethylsilyl-7-O-f3-Q-acetyl-2.4.6- trideoxy-α-L-threo-hexopyranosyl)adriamycinone <compound 121

A solution of glycosyl chloride (compound 7) in dichloro- methane (10 ml) prepared by hydrochlorination of hexenitol (295 mg, 1.89 mmol) was added to a suspension of 14-O-tert-butyldimethylsilyaldriamycinone (compound 11, 500 mg, 0.95 mmol), mercuric bromide (50 mg) , yellow mercuric oxide (750 mg) , and 4A molecular sieves (3.0 g) in dichloromethane (50 ml) . (Compound 11 can be prepared from daunomycinone (Compound 8) using generally the same procedure described by Horton et al., J. Antibiotics 37(8), 853-858 (1984), and by Priebe et al. in U.S. patent application serial no. 212,355, filed on June 27, 1988, which is incorporated herein by reference.) The resulting mixture was actively stirred at room temperature until disappearance of the substrate (60 minutes) . The salts were filtered off and the filtrate was diluted with dichloromethane (150 ml) and extracted with 10% aqueous potassium iodide (50 ml x 2) and water (50 ml x 2) . The organic extract was dried over sodium sulfate and evaporated. The residue was then chromatographed on silica gel (toluene-acetone 50:1) to give compound 12 (331 g, 51%) . First crystallization of TLC pure fractions (dichloromethane-hexane) gave analytically pure compound 12 (133 mg) ; mp 175-180°C; IRmax(KBr) 3487(OH), 1733(CO), 1617 and 1580(H-bonded quinone) , 1412, 1366, 1285 (SiMe) , 1109, 991, 978 and 838 cm^-1(CSi) . Analysis: Calculated for C₃₅H₄₄0₁₂Si (684.78):

C 61.39 H 6.48 Found: C 61.31 H 6.50

14-0-tert-Butyldimethylsilyl-7-0- ( 2 .4.6-trideoxγ-α-L- threo-hexopyranosvDadriamycinone (compound 13)

To a solution of compound 12 (150 mg, 0.22 mmol) in methanol (10 ml) was added with stirring 0.5 M sodium ethoxide in methanol (0.5 ml). After 40 minutes the reaction was terminated by adding dry ice. The mixture was then diluted with dichloromethane (100 ml) , washed with water (50 ml x 33), dried over sodium sulfate, filtered and evaporated. TLC showed one spot having Rf 0.19 (toluene-acetone, 8:1). The product was then crystallized from dichloromethane and hexane to give analytically pure compound 13; yield 101 mg (73%); mp 220°C; IRmax(KBr) 3475 (OH), 1732 (CO), 1618 and 1580 (H- bonded quinone) , 1458, 1442, 1430, 1412, 1285(Si e), 1207, 1107, 993, 976, 950, and 838 cm^_1(SCi).

Analysis: Calculated for C₃₃H₄₂0₁₁Si (642.76):

C 61.66 H 6.59 Found: C 61.58 H 6.59

7-0-(2.4.6-Trideoxy-α-L-threo-hexopryanosyl)adriamycinone (compound 14: 3'-deamino-3'-hvdroxy-esorubicin)

To a solution of compound 13 (100 mg, 0.1555 mmol) in oxolane (10 ml), dichloromethane (4 ml), and pyridine (1.0 ml) was added with stirring tetrabutylammonium fluoride (0.3 ml of a 1 M solu- tion in oxolane). After completion of the reaction (TLC, toluene-acetone, 3:1 Rf 0.25) in 30 minutes, the mixture was diluted with dichloromethane (150 ml) and water (50 ml x 3) . The organic layer was dried (sodium sulfate) and the residue after evaporation was purified by dissolving in dichloromethane and precipi- tated by addition of ethyl ether. The solid was washed with ether and dried to afford pure compound 19; yield 61 mg (74.4%) mp 160-163°C; IRmax(KBr) 3457(OH), 1720 (CO), 1618 and 1578 (H-bonded quinone), 1438, 1410, 1208, 1181, 1122, 1019, and 993 cm^"1.

The antitumor activity of compound 14 was tested against L-1210 murine leukemia in vivo. L-1210 cells (1 million) were inoculated intraperitoneally on day 0 to BDF1 mice. Groups of 6 mice each were used. Results were expressed as % T/C (median survival of treated animals: median survival of control animals x 100). Results obtained are shown below:

Drug Dose %T/C

(mg/kg)

Compound 14 12.5 180

25.0 490

50.0* 350

* The highest tested dose.

Compositions in accordance with the present invention can include a pharmaceutically effective amount of one or more of the novel antibiotic compounds and a pharmaceuti- cally acceptable carrier. The compositions can also contain solubility-enhancing agents such as DMSO or-the commercial surfactants Tween 20, Tween 80, Cremophor,. or Klucel. Alternatively, the active compounds might be formulated in a fatty emulsion, encapsulated in liposomes or polymeric drug carriers. Methods in accordance with the present invention comprise administering to a host an effective amount of the compounds or compositions described above. The administering step is preferably parenteral and by intravenous, intraarterial, intramuscular, intralymphatic, intraperitoneal, subcutaneous, intrapleural or intrathecal injection or by topical application or oral dosage. Such administration is preferably repeated on a timed schedule until tumor regression or disappearance has been achieved, and may be used in conjunction with other forms of tumor therapy such as surgery or chemotherapy with different agents.

The description and examples given in this patent are intended to illustrate the present invention. They are not intended to be an exhaustive list of all possible specific embodiments of the present invention. Those skilled in the art will recognize that modifications could be made to the specific embodiments listed here which would still be within the scope of the present invention.

Claims

CLAIMS :

1. A compound which has the formula

where R is selected from the group consisting of H, OCH₃, OH, and F;

2 R is selected from the group consisting of H_r OH, and carboxy having the formula COOR ;

R 3 is selected from the group consisting of COCH-R7 and

7_^ ^Δ

R 4 is selected from the group consisting of H, aliphatic and aromatic hydrocarbons having 1 to 10 carbons, and acyl o groups having the formula COR ;

R is selected from the group consisting of CH₃ and CH₂OH;

R is selected from the group consisting of H and aliphatic hydrocarbons having 1 to 6 carbons;

R is selected from the group consisting of H, OH, and g OR y

Q

R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons; 9 ,

R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons and COR ; and

R is selected from the group consisting of aliphatic hydrocarbons having 1 to 18 carbons.

2. The compound of clai.n 1, where R 1 is 0CH₃ and R2 i.s

H.

4 3. The compound of claim 2, where R is H

4. The compound of claim 2, where R 5 i.s CH,

5. The compound of claim 2, where R is selected from the consisting of H and OH.

6. A compound which has the formula:

7. A compound which has the formula:

8. A composition including an effective amount of a compound which has the formula:

and a pharmaceutically acceptable carrier.

where R is selected from the group consisting of H, OCH,, OH, and F;

R is selected from the group consisting of H, OH, and carboxy having the formula COOR ;

R 3 is selected from the group consisting of COC-H₂R7 and

CH₂CH₂R⁷; 4 R is selected from the group consisting of H, aliphatic and aromatic hydrocarbons having 1 to 10 carbons, and acyl

Q groups having the formula COR ;

R is selected from the group consisting of CH₃ and CH₂OH;

R is selected from the group consisting of H, OH, and g OR ;

R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons;

9 R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons and COR ; and

9. The composition of claim 8, where R 1 is ^"OCH₃ and R2 is H.

10. The composition of claim 9, where R 4 i.s H,

5 .

11. The composition of claim 9, where R is CH₃.

12. The composition of claim 9, where R is selected from the group consisting of H and OH.

13. A composition including an effective amount of a compound which has the formula:

and a pharmaceutically acceptable carrier.

14. A composition including-an effective amount of a compound which has the formula:

and a pharmaceutically acceptable carrier

15. A method of inhibiting neoplastic cell growth in a mammal, including the step of administering to a mammal an effective amount of a compound having the formula:

where R is selected from the group consisting of H, OCH-, OH, and F;

2

7 is selected from the group consisting of COCH-R and

CH₂CH₂R' ;

4 R is selected from the group consisting of H, aliphatic and aromatic hydrocarbons having 1 to 10 carbons, and acyl groups having the formula COR ;

R is selected from the group consisting of CH₃ and CH₂OH;

R is selected from the group consisting of H, OH, and OR³;

R .8 i.s selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons; 9 R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons and COR ; and

16. The method of claim 15, where Rl is OCH₃ and R2 is H,

17. The method of claim 16, where R is H.

18. The method of claim 16, where R is CH 3'

19. The method of claim 16, where R is selected from the group consisting of H and OH.

20. A method of inhibiting neoplastic cell growth in a mammal, including the step of administering to a mammal an effective amount of a compound having the formula:

21. A method of inhibiting neoplastic cell growth in a mammal, including the step of administering to a mammal an effective amount of a compound having the formula:

22. A method of inhibiting lymphoid leukemia cell growth in a mammal, including the step of the step of administer¬ ing to a mammal an effective amount of a compound having -20 the formula:

where R is selected from the group consisting of H, OCH₃, OH, and F;

35

R is selected from the group consisting of COCH₂R and 40 CH₂CH₂R⁷; 4 R is selected from the group consisting of H, aliphatic and aromatic hydrocarbons having 1 to 10 carbons, and acyl o groups having the formula COR ;

R is selected from the group consisting of CH- and CH₂0H;

R is selected from the group consisting of H, OH, and g OR*;

Q

R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons; g R is selected from the group consisting of aliphatic hydrocarbons having 1 to 6 carbons and COR ; ar 3

23. A method of inhibiting lymphoid leukemia cell growth in a mammal, including the step of administering to a mammal an effective amount of a compound having the formula:

24. A method of inhibiting lymphoid leukemia cell growth in a mammal, including the step of administering to a mammal an effective amount of a compound having the formula: