CA2228595C - Novel water-soluble c-ring analogues of 20 (s)-camptothecin - Google Patents

Abstract

Novel water-soluble C-ring analogues of 20(S)-camptothecin having general formula (1). All the compounds of formula (I) are prepared from the compounds of general formula (2) having 20(S)-chiral carbo n where R1 to R5 have the meaning given above. The compounds of formula (1) possess potent anti-cancer and anti-viral propertie s. The invention also provides an alternate process for the preparation of known C-5 substituted compounds of formula (1).

Description

NOVEL WATER SOLUBLE C-RING ANALOGUES
OF 20(S)-CAMPTOTHECIN
The present invention relates to novel water soluble C-ring analogues of 20(S)-Camptothecin having the general formula Z.

In the above formula l, R1, R2, R3, R4 are independently the same or different and represent hydrogen, hydroxy, aryloxy, lower alkoxy, lower alkanoyl, vitro, cyano, halo, carboxy, carbonyloxy, 3.0 amino, substituted amino, lower alkyl, substituted lower alkyl, or R2, R3 together represent -O-(CH2)n-O-where n=1 or 2; R5 represents hydrogen, lower alkyl, substituted lower alkyl, lower aralkyl, hydroxymethyl, carboxymethyl, aminomethyl, substituted aminomethyl Z5 where the amino group may be mono or disubstituted a.n ' which both substituents are independent or combined together to form a cyclic ring system of a total of 5-6 ' atoms containing carbon and optionally one or two heteroatoms selected from oxygen, nitrogen or sulfur;

and R6 represents hydrogen, phenyl or benzyl where the phenyl group may be unsubstituted or substituted with mono, di or trisubstituents which may be selected from halogen, hydroxy, lower alkoxy, cyano, carboxyl, vitro, amino or substituted amino, lower alkyl, substituted lower alkyl; cycloalkyl or cycloalkyl lower alkyl where the cyclic ring is in the range of 3 membered to 7 membered ring system containing all carbon atoms; lower alkyl groups substituted with heterocyclic rings where the heterocyclic ring system has a total of 3 to 7 atoms, the heterocyclic rings containing carbon with at least one heteroatom such as oxygen, nitrogen or sulfur; lower alkanoyl; benzoyl where the phenyl group can be unsubstituted or substituted; lower alkenyl;
lower alkyl; substituted lower alkyl, substituted lower alkenyl or substituted lower alkanoyl where the substituents can be halogen, hydroxy, lower alkoxy, aryloxy, thio, thioalkyl, thioaryl, aryl, heteroaryl, carboxy, cyano, vitro, amido or amino in which the amino group can be unsubstituted or mono, or disubstituted in which both substituents are independent or combined together to form 5 or 6 membered cyclic ring system containing carbon, and optionally contain one or two heteroatoms selected from oxygen, nitrogen or sulfur, the total number of atoms in the cyclic ring system is 5 or 6; with the proviso that (i) when R1 is methoxy group, R6 is not hydrogen or lower alkyl group; (ii) when R2 is hydroxy, lower alkoxy, thioalkyl, vitro, amino. alkylamino, acylamino or halogen, R6 is not hydrogen or lower alkyl group;
(iii) when R5 is lower alkyl, lower aralkyl, CH20H, COON, COOMe or CH20R" where R" represents lower alkyl or acyl group, R6 is not hydrogen or lower alkyl group;
(iv) when R1 is methoxy group, R2 is hydroxy, lower alkoxy, thioalkyl, vitro, amino, alkylamino, acylamino, or halogen, R5 is lower alkyl, lower aralkyl, CH20H, COOH, COOMe or CH20R" where R°' represents lower alkyl or acyl group, R6 is not hydrogen or lower alkyl group;
(v) when R1 through R~ represent hydrogen, R6 is not hydrogen or lower alkyl group.
All these compounds of the formula 1 are prepared from the compounds of the general formula 2 having 20(S)-chiral center, A ' B,1 c\ 16 O
' p t7 R4 1 13. .,.. 20 where RZ to R~ have the meaning described above.
Camptothecin having the formula 3, is an alkaloid with strong antitumor activity, / O

and was isolated from Camptotheca acuminata by Wall and co-workers in 1966. However, its development as a potential drug for cancer treatment had been abandoned due to unacceptable side effects on humans and due to its low water solubility as well as high toxicity problems. Since the discovery of its mechanism of action as an inhibitor of topoisomerise 2 by Liu and co-workers in 1985 [L. F. Liu, et al., J. Biol. Chem., 260, 14873 (1985)], the research interest on camptothecin has once again taken momentum.
To overcome this problem of low water solubility and high toxicity of camptothecin, over the last 30 years, several research groups all over the f~ lB~ C N 6 12 N E o tHO~

world have prepared and investigated a number of camptothecin analogues involving the modification of rings A-E'or the introduction of a variety of substituents on all the five rings of camptothecin of the formula 3 [M. E. Wall et al. , J. Med. Chew. , 36, 2689 (1993); R. P. Hertzberg et al., J. Med. Chem., 715 (1989): S. W. Sawada et al., Chew. Pharm. Bull., 42(2), 310 (1993)]. Among the various camptothecin analogues prepared to date, only two of them namely, CPT-11 having the formula 4 [Chew. Pharm. Bull., 39, 1446 (1991) ] , a _ o .HCl,3H20 \
H
- o topotecan of the fox~ula 5 [J. Med. Chem., 34, 98(1991)]
lSMe2 .HCI
- ~, r w w i'.
H
r-=
S
were introduced as anti-cancer drugs in the market recently. Another compound namely, 9-aminocamptothecin of the formula 6 [J. Med. Chem., 29, 2358 (1986)], ~. , v v w' HO
O
G

WO 97/46563 ~CTliJS97/06962 is currently undergoing extensive clinical trials. The extensively studied Structure Activity Relationship (SAR) on camptothecin of the formula 3 [M. E. Wall et al., J. 3~Ied. Chem., 36, 2689 (1993)7 has revealed that 20 (S) -«-hydroxy-b-lactone (E-ring) moiety in camptothecin is essential for its activity. However, according to recent reports by Ejima et al., replacement of hydroxyl group with an amino group at C-20 position leading to a compound such as 7-ethyl-10-methoxycamptothecin derivative of the formula 7 [A.
Ejima et al., Chem. Pharm. Bull., 40(3), 683 (1992)], Me r ~. r~
i o HO
zo ~o %" o 7 $
exhibited an increased in vivo antitumor activity than 20(RS)-camptothecin of the formula 8. Also in another report (Lawrence Snyder et al., J. Org. Chem., 59, 7033 (1994)x, the I8-noranhydrocamptothecin analogue of the formula 9, .._ exhibited potent camptothecin lilte inhibition of ' topoisomerase I activity. Both these reports are contrary to the assumption that 20(S)-«-hydroxy functionality in camptothecin is an essential feature for its biological activity.

WO 97/46563 fCT/US97106962 _ g _ Based on the structure-activity results obtained for the camptothecin analogues prepared in the literature, it was established that the modification of substituents at C-9 and C-7 position of camptothecin of the formula 3 plays an important role in the enhancement of anticancer activity by imparting stability to the E-ring lactone [T. G. Burke et al., J.
Med. Chew. 37, 40 (1994)]. It has also been recognized that the open form of the lactone moiety, namely, 't~ze Carboxylate form' is less effective therapeutically than the closed 'Lactone form' [Hertzberg et al., J.
Med. Chem., 32, 715(1989): J. M. Covey, C. Jaxel et al., Cancer Research., 49, 5016 (1989); Giovanella et al., Cancer Research., 51, 3052 (1991)3. The recent studies by T. G. Burke et al., on the stability of 'closed lactone form' of various camptothecin analogues in the presence of protein called 'Human Serum Albumin' (HSA) indicated that the compounds such as CPT-11 of the formula 4 and 7-ethyl-10-hydroxycamptothecin (SN-38) of the formula 7a 7a and Topotecan of the formula 5, in the presence of HSA
at 37°C., exhibited a higher percentage (o) of lactone form at equilibrium than 20(S) camptothecin of the formula 3 and 9-aminocamptothecin of the formula 6 [T.
G. Burke and Zihou Mi., J. Med. Chem., 37, 40 (1994);
ibid., Biochemistry., 33, 12540 (1994).]. Based on these studies, it was recognized that the understanding of the factors influencing the lactone-carboxylate equilibrium of camptothecin analogues became an important determinant in the design of novel and - 7 _ therapeutically efficacious drug candidates a.n the camptothecin series.
Although the modification of substituents on rings A and B of camptothecin was taken up at a rapid pace to generate novel CPT analogues, ring 'C' analogues of camptothecins were limited presumably.
because of the research work carried out by Sawada et al., which claimed that the substituents at C-5 position of camptothecin has resulted in the reduction of anti-tumor activity of camptothecins and produced inactive analogues [Sawada S. et al., Chem. Pharm.
Bull., 39(10), 2574 (1991)]. The C-5 substituted camptothecins claimed by Sawada et al., (JP 58,154,584;
US 4,513,138; US 4,473,692; US 4,545,880; US 4,339,282) have the structural formula 10, IO
where R represents hydroxy, lower alkyl, lower alkoxy, acyloxy groups, R1 represents hydrogen, methoxy at 9th position; hydrogen, hydroxy, lower alkoxy, acyloxy, SH, thioalkyl, thioacyl, vitro, amino, alkylamino, acylamino and halogen groups at 10th position and R2 represents hydrogen, lower alkyl, lower aralkyl, CH20H, COOH, COOMe, CH20R' where R' represents lower alkyl or acyl group.
The recent findings by K. H. Lee et al.,[Bio.
Org. Med. Chem. Lett., 5(1), 77 (1995)] which includes the preparation of 5-hydroxymethyl camptothecin by the reaction of formaldehyde in N,N-dimethylformamide and 4-piperidinopiperidine on 20(S)-camptothecin, has revealed the reduced anti-tumor activity of these _ g _ compounds. Also, Danishefsky et al., prepared some of the C-5 substituted 20(RS)-camptothecin derivatives by a totally synthetic approach [US 5,391,745 and US ' 5, 446, 047] .
However, the synthetically prepared 5-substituted camptothecin derivative of the formula 11 [Terasawa et al., Heterocycles, 38, 81 (1994)] claimed to have anti-tumor activity comparable to that of 20(S)-camptothecin.
is Keeping all these factors in mind, we focused our research studies on 20(S)-camptothecin aimed at the design of novel camptothecin analogues which can exhibit improved water solubility and improved stability of lactone form in solution. We identified a oxidative reaction in alcoholic solvents for this purpose. The resultant findings have culminated into the discovery of a novel synthetic transformation which can introduce a variety of alkoxy groups at C-5 position of 20(S)-camptothecins. Functional group transformation of these 5-alkoxy camptothecins produced a wide variety of novel C-5 substituted 20(S)-camptothecin analogues of the formula 14, t2 ~ g D 97 in which X represents NH or NR and CHZ or CHR and R6 has the meaning described above, which is the subject matter of our U.S. Patents 6,214,836 and 5,972,955.
Hence, the discovery led to a facile and versatile semi-synthetic methodology by which virtually every camptothecin derivative known in the literature can be transformed into a variety of C-5 substituted camptothecin analogues.
Therefore, the present invention provides a novel process for the preparation of various C-5 substituted 20 (S)-camptothecin derivatives of the formula where R6 has the meaning described above. Also, by virtue of the present invention, a second chiral center at C-5 position was introduced into the camptothecins of the general formula 2 without disturbing the existing 20-hydroxyl group, C-20 (S) chiral center.
Furthermore, the vast variety of substituents represented by OR6 at the C-5 carbon of (S)-camptothecins of the formula 1 led to compounds with improved water solubility ranging from lmg to l Omg per ml. All of the compounds prepared by the 15 present invention exhibited significant in vitro anti-tumor activity against a wide range of human tumor cell lines.
Detailed Description of the Invention The present invention particularly provides C-5-O-substituted water soluble analogues of 20 (S)-Camptothecin having the formula 1 f i R 1 R 5 ,.~s where R', RZ, R3 , R4, RS and R6 have the meaning WO 97/46563 I'CT/CTS97/06962 described above. Throughout the present invention, the terms representing RI through R6 in these compounds have the following definitions. , The term 'lower alkyl' denotes a univalent, branched or straight hydrocarbon chain containing 1 to 8 carbon atoms. Representative of the alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, sec. butyl tert.butyl pentyl, iso pentyl tert.pentyl, hexyl, isohexyl and octyl.
The term 'lower alkenyl' represents a branched or straight hydrocarbon chain having sp or sp2 carbon centers containing 1 to 8 carbon atoms.
Representative of the alkenyl groups are vinyl, propenyl, butenyl pentenyl, isopropenyl, isobutenyl, proparginyl, hexenyl and octenyl.
The term 'halogen' or 'halo' represents chlorine, bromine or fluorine. The term 'haloalkyl' denotes alkyl groups substituted with halogens, preferably fluorine, bromine or chlorine.
Representative of the haloalkyl groups are chloroethyl, bromopropyl, fluoroethyl, trifluoroethyl, trichloroethyl and trifluorobutyl.
The term 'lower alkoxy' denotes lower alkyl groups as defined above attached via oxygen linkage to the rest of the molecule. Representative of those groups are methoxy, ethoxy, isopropoxy, tert.butoxy, hexoxy, heptoxy and octoxy.
The term 'lower alkanoyl' denotes lower alkyl or alkenyl groups as defined above attached via a carbonyl group to the rest of the molecule.
Representative of those groups are acetyl, propionyl, propenoyl, crotanoyl, butanoyl, pentanoyl and ' isopentanoyl.
The term 'aminoalkyl' represents the lower ' alkyl groups as defined above substituted with amino groups. Representative of the aminoalkyl groups are 2-aminopropyl, 4-aminobutyl, 5-aminopentyl. Amino groups WO 97/46563 PCTlLTS97/06962 may also be mono or disubstituted and the representative of these substituted amino groups are . dimethylamino, diethylamino, dibenzylamino, ethylisopropylamino, pyrrolidino, piperidino, . 5 morphilino or piperizino.
The term 'heteroatom' refers to oxygen, nitrogen or sulfur. The term 'aryl or heteroaryl' represents the groups of aromatic nature having 5 or 6 membered rings which may be selected from phenyl, biphenyl, naphthyl, pyridyl, quinoline, isoquinoline, indole, pyroll, furan, benzofuran, thiophene, pyramidine, piperizine, thiosolidine or imidazole.
The term 'substituted phenyl' group used in the present invention refers to those substituents which can be selected from the groups such as hydroxyl, lower alkyl, haloalkyl, phenyl, benzyl, halogen, lower alkoxy, thioalkoxy, benzyloxy, carboxyl, cyano, vitro, amido, amino, and alkylamino. Examples of such groups are 4-hydroxyphenyl, 3-methoxyphenyl, 4-fluorophenyl, 4-trifluoromethylphenyl, N,N-dimethylaminophenyl, and 4-carbomethoxyphenyl.
The term 'substituted alkyl' group used in the present invention refers to those substituents which can be selected from the groups such as hydroxyl, alkyl, haloalkyl, phenyl, benzyl, halogen, alkoxy, thioalkoxy, benzyloxy, carboxyl, carbonyloxy, cyano, vitro, amido. amino, and alkylamino. Examples of such groups are fluoroethyl, chloropropyl, hydroxyethyl, methoxypropyl, N,N-diethylaminoethyl, N-benzoylaminopropyl, trifluoroethoxyethyl, phenoxyethyl, carbomethoxyethyl, (p-fluorobenzoyloxy)ethyl, ' aminopropyl, and 2-thioethyl.
The term 'substituted amino' group used in the present invention refers to those substituents which can be selected from the groups such as hydroxyl, alkyl, haloalkyl, benzyl, benzoyl, alkoxy, carboxyl, amido, amino, and alkylamino. Examples of such groups W~ 97146563 PCTIUS97/06962 are N,N-diethylamino, N-benzoylamino, N-methoxyamino, N-carboethoxyamino, and N-chloroethylamino groups.
Also, both the substituents on the amino group can be combined together to form 5 or 6-membered cyclic ring system represented by pyrrolidino, piperidino, piperizino, morphilino, imidazolino, or thiazolidino.
According to the present invention there is provided a process for the preparation of the compounds of the general formula 1, ._ N O

t E
t-t0 wherein R1, R2, R3, R4 are independently the same or different and represent hydrogen, hydroxy, aryloxy, lower alkoxy, lower alkanoyl, vitro, cyano, halo, carboxy, carbonyloxy, amino, substituted amino, lower alkyl, substituted lower alkyl or R2, R3 together represent -O-(CH2)n-O- where n=1 or 2; RS represents hydrogen, lower alkyl, substituted lower alkyl, lower aralkyl, hydroxymethyl, carboxymethyl, aminomethyl, substituted aminomethyl where the amino group may be mono or disubstituted in which both substituents are independent or combined together to form a cyclic ring system of a total of 5-6 atoms containing carbon and optionally one or two heteroatoms selected from oxygen, nitrogen or sulfur; and R6 represents hydrogen; phenyl or benzyl where the phenyl group may be unsubstituted ' or substituted with mono, di or trisubstituents which may be selected from halogen, hydroxy, lower alkoxy, cyano, carboxyl, vitro, amino or substituted amino, lower alkyl, substituted lower alkyl; cycloalkyl or cycloalkyl lower alkyl where the cyclic ring is in the range of 3 membered to 7 membered ring system containing all carbon atoms; lower alkyl groups substituted with heterocyclic rings where the heterocyclic ring system has a total of 3 to 7 atoms, the ring system contains carbon with at least one heteroatom such as oxygen, nitrogen or sulfur; lower alkanoyl; benzoyl where the phenyl group can be unsubstituted or substituted; lower alkenyl; lower alkyl; substituted lower alkyl, substituted lower alkenyl or substituted lower alkanoyl where the substituents can be halogen, hydroxy, lower alkoxy, aryloxy, thio, thioalkyl, thioaryl, aryl or heteroaryl, carboxy, cyano, vitro, amido or amino in which the amino group can be unsubstituted or mono, or disubstituted a.n which both substituents are independent or combined together to form 5 or 6 membered cyclic ring system containing carbon, and optionally contains one or two heteroatoms selected from oxygen, nitrogen or sulfur, the total number of atoms a.n the cyclic ring system being 5 or 6; with the proviso that (i) when Rl a.s methoxy group, R6 is not hydrogen or lower alkyl group; (ii) when R2 is hydroxy, lower alkoxy, thioalkyl, vitro, amino, alkylamino, acylamino, and halogen, R6 is not hydrogen or lower alkyl group ; (iii) when R5 is lower alkyl, lower aralkyl, CHZOH, COOH, COOMe, or CH2OR" where R"
represents lower alkyl or acyl group, R6 is not hydrogen or lower alkyl group (iv) when R1 is methoxy group, R2 is hydroxy, lower alkoxy, thioalkyl, vitro, amino, alkylamino, acylamino, or halogen, R5 is lower alkyl, lower aralkyl, CH20H, COOH, COOMe or CH20R"
~ where R" represents lower alkyl or acyl group, R6 is not hydrogen or lower alkyl group; (v) when R1 through ~ R5 represent hydrogen, R6 is not hydrogen or lower alkyl group, which comprises, (i) reacting the compounds of the formula 2, WO 97/46563 fCT/IJS97106962 5 _ O
0 0 ~ ~N
,2 D 17 ... w ~ 3 E .., where R1 to RS have the meaning described above, in the presence of an acid and an oxidizing agent which is a.
ferric salt, with a compound having the formula R6-OH
where R6 represents lower alkyl, lower alkenyl, (C3-5 C7)cycloalkyl, haloalkyl or hydroxyalkyl, to obtain compounds of the formula 12 and compounds of the formula 13, C O ~ c O

I2 4 ~ 3 E R 12 3 ' D ~7 R HO 20 O R4 _20 t-10 O _ O
xs wherein R1, R2, R3, R4, R5 have the meaning given above, (fi) separating the compounds of the formulae 12 and 13 prepared in the step (i), by conventional methods, (iii) hydrolyzing the compounds of the formula 12, by conventional methods, to obtain additional amounts of the compounds of the formula 13, I5 (iv) reacting the compound of the formula 13, ' in the presence of an acid, with a compound having the formula R6-OH to obtain compounds of the formula 1, ' _ l5 _ C ~H O
t7 12 '"
R4 t HO 20 <
O
x where RI, R2, R3, R4 and R5 have the meaning described above and R6 is as defined above.
According to another feature of the present invention there is provided an alternate process for the preparation of known C-5 substituted compounds of the formula 1, 9' R

O

1 R = 3 ''D~ E1 <
EiO
x wherein R6 represents hydrogen or lower alkyl, R1 represents hydrogen or methoxy; R2 represents hydrogen, hydroxy, lower alkoxy, acyloxy, thioalkyl, SH, thioacyl, ~ ~10 vitro, amino, alkylamino, acylamino and halogen; R3 and R4 are hydrogen and R5 represents hydrogen, lower alkyl, lower aralkyl, CH20H, COOH, COOMe or CH20R' where R' represents lower alkyl or acyl group which comprises, (i) reacting the compounds of the formula 2, t 5 . R 9 ~ 5 O
C IS
R 1~~ O t7 l 3 E \
Et HO 20 O
O

WO 97/46563 PCTlLTS97/06962 where R1 to R5 have the meaning described above, in the presence of an acid and an oxidizing agent such as ferric salt, with a compound having the formula R6-OH
where R6 represents lower alkyl groups, to obtain compounds of the formula 12 and compounds of the formula 13, i 5 ~n~
R
O
C \(~
i2 3 o t7 R Z2 3 O 17 a4 ~ _~ n _w Z
x2 wherein RZ, R2, R3, R4 and R5 have the meaning given above, (ii) separating the compounds of the formulae 12 and 13 prepared in the step (i), by conventional methods, (iii) hydrolyzing the compounds of the formula 12, by conventional methods, to obtain additional amounts of the compounds of the formula 13, (iv) reacting the compound of the formula 13, in the presence of an acid, with a compound having the formula R6-OH to obtain compounds of the formula 1, R6 represents lower alkyl groups, R1 represents hydrogen or methoxy, R2 represents hydrogen or hydroxy, lower alkoxy, acyloxy, SH, thioalkyl, thioacyl, vitro, amino, alkylamino, acylamino and halogens R3 and R4 are hydrogen and R~ represents hydrogen, lower alkyl, lower aralkyl CHZOH, COOH, COOMe or CH20R' where R' represents lower alkyl or acyl group.
The methodology developed and described in the present invention has generated a new chiral center at C-S position in the compounds of formula 2 without disturbing the integrity of 20(S)-«-hydroxy E-ring lactone moiety. The process developed constitutes a novel, facile and versatile semi-synthetic method for the preparation of C-5 substituted known and novel camptothecin derivatives of the formula 1, starting from the compounds of formula 2. The compounds of the formula 1 prepared by the process of the present invention thus represents diastereomers containing the newly created C-5 chiral center. Indeed, the compounds of the general formula 1 are isolated as a mixture of 20(S).5(R) and 20(S),5(S) diastereomers. However, by the application of conventional analytical techniques, the two diastereomers have also been separated into their single optically pure entities.
In general, all of compounds of the formula 1 where R1, R2, R3, R4, R5 and R6 have the meaning described above. may be synthesized starting from the compounds of the formula 2 by the process described above and can be illustrated with the examples described in the Examples Section. The preparation of the compounds of the formula 12, where R1, R2, R3, R4, R5 and R6 have the meaning given above, from the compounds of the formula 2 as mentioned in the step (i), is a novel transformation in which a direct introduction of various types of al3coxy substituents at C-5 position has been achieved.
The A ring or A/B ring substituted 20(S)-camptothecin derivatives of the general formula 2 where R1, R2, R3, R4 and R5 have the meaning described above, used as starting materials in the present invention are widely known and prepared according to the prior art documented in the literature. For example, 7-ethylcamptothecin, 10-hydroxycamptothecin, 9-nitrocamptothecin, 12-nitrocamptothecin, 10-hydroxy-7-ethylcamptothecin (SN-38), 9-amino-camptothecin, 9-methoxycamptothecin, 9-hydroxycamptothecin, 9-methoxy-7-ethylcamptothecin, 9-hydroxy-7-ethylcamptothecin, 10,11-methylenedioxycamptothecin, 10,11-ethylenedioxycamptothecin, 10-hydroxy-9-(N,N-dimethylaminomethyl)camptothecin were prepared according to the known literature methods CT. R.
Govindachari et al. , Ind. J. Chew. 10 {B) , 453 {1972) ; S.
Sawada et al, Chew. Ph.arm. Bull, 39(10) 2574 (1991), ibid., 39(12), 3183 (1991); US patent no. 4,604,463;
and US 4,545,880; Jaffery L. Wood et. al., J. Org.
Chem., 60 5739 (1995)] and used as starting materials for the preparation of novel C-ring substituted 20(S)-camptothecin analogues of the general formula 1 described in the present invention.
For example, compounds of the formula 12 where R1, R2, R3, R'~, R5 and R6 are independently the same or different and have the meaning given above, can be prepared, as mentioned in the step (i), by the reaction of the compounds of the formula 2 with the compounds having the formula R6-OH where R6 represents hydrogen, lower alkyl, lower alkenyl, haloalkyl, hydroxyalkyl, (C3-C7) cycloalkyl, in the presence of a strong acid and a ferric salt. The acids used in this reaction can be selected from perchloric acid, hydrochloric acid, nitric acid, sulfuric acid or Lewis acids such as boron-trifluoride, zinc chloride, tinchloride, titanium tetrachloride. The ferric salt used in the above reaction can be chosen from ferric nitrate, ferric ammonium sulfate, ferric chloride. In general, the above reaction may be affected at a temperature in the range of 40-150°C., preferably 60 'to 120°C.
In step (ii) of the s~rocess of tha r~rPr~~."t --_ r__....»-... -invention, to separate the mixture of compounds of formulas 12 and 13 as prepared a.n the step (i) the mix-ture is subjected to preferably either crystallization or column chromatography technique using silica gel.
The solvent mixtures used in the above mentioned methods may contain a combination of the organic solvents such as chloroform, ethyl acetate, methanol, ethanol, ether, acetone and hexane.
The compounds of the formula 13 can also be obtained in step .(iii) of the process of the present invention, by treating the compounds of the formula 12 with acids a.n combination with water at a temperature in the range of 40-120°C. The acids used for this purpose may be selected from hydrochloric acid, hydrobromic acid, sulfuric acid, p-toluenesulfonic acid, acetic acid and perchloric acid. The solvents used in the reaction may be methanol, ethanol, butanol, isopropanol or 1,4-dioxane.
In step (iv) of the process of the present invention, compounds of the formula 13 were reacted with compounds of the formula R6-OH where R6 has the meaning described above, in the presence of an acid medium at a temperature in the range of 20 to 140°C. to furnish the compounds of the formula 1. The acids used in the reaction may be selected from sulfuric acid, hydrochloric acid, acetic acid, p-toluenesulfonic acid, pyridinium-p-toluenesulfonic acid, camphorsulfonic acid, methanesulfonic acid, perchloric acid or Lewis acids such as titanium tetrachloride, BF3-etherate and zinc chloride. The solvents used in the reaction may be selected from hexane, benzene, toluene, xylene, chloroform, carbon tetrachloride, dichloroethane, dichloromethane and 1,4-dioxane.
Thus, the present invention a.s of particular significance in developing C-5-substituted 20(S)-camptothecin derivatives as a new class of C-ring modified camptothecin analogues which are useful as anti-tumor and/or anti-viral agents. The present invention is also of particular significance as the process developed and described here a.s highly < versatile and amenable for large scale preparation of these camptothecin derivatives having the general formula 1.
The methodology developed and described a.n the present invention will provide access to a wide variety of substituted C-ring analogues having diverse substituents on rings A
and B
of 20(S)-campto--thecin.
Some of the preferred compounds are those where is vitro, amino, aminoalkyl, hydroxy, methoxy;

R2 is hydroxy, carbonyloxy, halo; R2, R3 combined together to represent methylenedioxy or ethylenedioxy;

R4 is hydrogen or vitro; R5 is ethyl, aminomethyl or substituted aminomethyl;

is 2'-hydroxyethyl, alkoxyethyl, chloroethyl, fluoroethyl, trifluoro-ethyl, or aminoethyl or aminopropyl where amino group may be dimethylamino, diethylamino, pyrollidino, piperidino, morphilino, piperizino, imidazolino;

Representative of the compounds of formula are:

1) 5-methoxy CPT*

2) 5-ethoxy CPT*

3) 5-butoxy CPT*

4) 5-chloroethoxy CPT*

5) 9-methoxy-5-ethoxy CPT

6) 9-hydroxy-5-ethoxy CPT

7) 10-hydroxy-5-ethoxy CPT*

8) 7-ethyl-5-ethoxy CPT*

9) 7-ethyl-5-hydroxy CPT*

10) 9-vitro-5-ethoxy CPT

11) 9-vitro-5-hydroxy CPT

12) 7-Ethyl-5-chloroethoxy CPT

13) 10-hydroxy-7-ethyl-5-ethoxy CPT*

14) 5-(2'-hydroxyethoxy) CPT

15) 7-ethyl-9-hydroxy-5-ethoxy CPT

16) 10-hydroxy-5-(2'-hydroxyethoxy) CPT

17) 7-ethyl-10-hydroxy-5-(2'hydroxyethoxy) CPT

18) 9-vitro-5-fluoroethoxy CPT

29) 9-vitro-5-trifluoroethoxy CPT

20) 10-hydroxy-5-trifluoroethoxy CPT

21) 7-ethyl-10-hydroxy-5-trifluoroethoxy CPT

22) 7-ethyl-5-pyrrolidinoethoxy CPT

23) 7-ethyl-5-dimethylaminopropoxy CPT

WO 9?/46563 PCT/US97/06962 24) 7-ethyl-10-hydroxy-5-fluoroethoxy CPT
25) 5-(2'-hydroxyethoxy)-7-ethyl CPT
26) 5- (2' -methoxyethoxy) CPT
where CPT refers to 20(S)-camptothecin S and ~ represents known compounds in the literature.
Most of the compounds prepared by the present invention have water solubility ranging from 1 mg to 10 mg per ml at 37°C. Table lA shows MTD in Swiss Albino mice, Lactone Stability in whole blood (after 3 hours), Solubility, Pharmacakinetics MTD, and In vitro activity after 1 hour exposure, for the compounds of Examples 11, 26 and 27.
The protocols used for conducting the experiments are:
l5 1. MTD in Swiss albino mice:
Each Swiss albino mouse is injected with a single does of the test compound on a day designated as Day l:
Doses that were tested are 400, 200, 100, 50, 25, 12.5, 8.3, 6.25 and 3.13 mg/kg body weight. The animals were observed for mortality and morbidity daily and the body weights of surviving animals were recorded on days 1, 5, 10 and 14. The maximum tolerated dose is defined as the dose at which the test compound did not exhibit any morbidity and body weight reduction more than 30~ as compared to day one. (As per the protocol followed by the U.S. National Cancer Institute) 2. Lactone Stability a.n whole blood:
2 ml of blood from a healthy volunteer was collected in a tube containing 40 Ea,l of heparin (572 IU) to prevent coagulation, 4 mM and 40 ~,cM working solutions of the drug in DMSO were prepared and added to aliquots of whole blood to give final concentration of 100 ~.M and 1 Ea.M respectively. The drug is incubated in whole blood at 37°C. and 20 /.cl samples are collected into 180 E.i.l of chilled methanol (-30°C.) at different time intervals (0, 1, 2 & 3 hrs). Vortex and they centrifuge at 11000 rpm for 3 min in a microcentrifuge at room temperature.
Dilute 100 E.cl of the supernatant with water to 300-500 _ /,cl depending upon the signal response (W for 100 ~M
concentration and fluoroscence for 1 /.cM concentration) of the compound. 200 Ecl of the diluted sample is injected on t'o the HPLC column previously equilibrated with the mobile phase. The area under peak corrsponding to lactone forms is measured. Zero time peak area is taken as 100, and the proportion of lactone peak area at different time points is calculated to determine the equilibrium lactone stability as obtained by consistent lactone proportion over two successive time points. (Biochemistry, 1994;
33:10325-10336 and J. Pharm. Sci., 1995; 84:518-519).
3. Pharmacokinetics at MTD:
A11 studies were carried out in Swiss albino mice in the weight range 35-40 g. The animals were fasted overnight prior to dosage of the drug and were fed 3 hours after dosing. The animals were dosed intra-peritonially as a solution in DMSO:water (50:50; v/v).
Blood samples were drawn from orbita3 sinus at 1, 2, 4, 6 and 8 hours after administration of the dose into heparinized tubes, centrifuged at 13000 RPM for 10 min.
Plasma samples were separated and analyzed by HPLC. To 50 /,cl of the sample, 100 /,cl of chilled acidified methanol was added and mixed to precipitate proteins.
The sample was centrifuged at 13,000 RPM for 10 min.
100 /.cl of supernatant was made up to 200 E,cl with methanol:wate_r (50:50; v/u) and 1_00_#,1 S"Fasin~eeted or,.
HPLC. Peak area of the drug was used for quantification. Calibration, control and recovery ' samples were prepared by spiking 50 ~.C1 of blank plasma with known amounts of the drug and processed in the ' same manner as the samples. (J. Natl. Cancer Inst.
1996; 88:817-824).

4. Solubility by HPLC method:
Excess of compound was soaked in 0.5 ml of 0.1M sodium acetate buffer at. pH 5.0 for 24 hours at room temperature. The solution was filtered through 0.45 micron PVDF syringe filter (Gelman Sciences). The filtrate was injected into HPLC at different volumes (10 & 20 ~e.l). Chromatograms were recorded. Responses recorded were extrapolated from the calibration curve and the solubility of the compound was calculated.
Med. Chem., 1995; 38: 400) 5. Solubility by routine method:
Compound was suspended in 5 ml of deionized water and heated to 37°C. for 10 min. Then, the solution was filtered and the filtrate was evaporated to drvinQ
using methanol and the solid residue was weighed.
6. In vitro activity after 1 hour exr~osure:
Grow the cells in 15 ml of Complete Medium (RPM1-1640 with 10% Fetal bovine serum and 0.2o NaHC03) for 3-5 days to obtain a cell number of 106 cells/flask. The medium is removed and the attached cells are washed with Phosphate Buffered Saline (PBS). 1 ml of 0.1~
Trypsin-EDTA is added and incubated for 5 min at 37°C.
Tap the flasks gently and add 5 ml of complete medium.
Remove cell suspension and centrifuge at 2000 rpm for 5 min. Discard the supernatant and suspend the pellet a.n 5 ml of complete medium. Count the cell number in a haemocytometer. Dilute the cell suspension to 10,000 cells/100 p,l in complete medium. Plate out 100 ~.cl of cell suspension in each of 96 well microtitre plate and incubate for 24 hrs at 37°C. and 5% C02. Terminate the reference blank (plated out separately) with 25 ~.r,l of 505 cold Trichloroacetic acid (TCA). Incubate for 1 hour at 4°C. Wash the plate (five times) with . deionized water. Air dry and preserve the plate at 4°C. for determination of TD value. Prepare suitable dilution of the test compound in complete medium and add 100 E,cl to each well to maintain the final concentration ranging between 10-4M and 10-8M. Incubate for 1 hr at 37°C. and 5o C02. Centrifuge the microtitre plate at 1000 rpm for 5 min. Remove the supernatant. wash the cells twice with 100 J,cl of PBS
to remove the traces of test compound. Add 200 ~r.l of complete medium to each well and incubate for 48 hours at 37°C. and 5~ C02. Terminate the cell growth with the addition of 50 Ecl of 50~ cold TCA. Incubate the plate for 1 hr at 4°C. Wash the plates with deionized water (five times) and air dry. Add 100 lcl of Sulforhodamine B solution (0.4~ in l~ acetic acid) to each well. Keep at room temperature for 15 minutes.
Wash (five times) with l~ acetic acid and air dry. Add 100 l.cl of lOmM Trizma base {Sigma) , shake gently on plate shaker for 15 minutes and read the optical density at 490 nm in Spectrophotometric plate reader.
(as per the protocol followed by the U.S. National Cancer Institute.

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WO 97!46563 PCTlLTS97/06962 Further, several compounds prepared in the present invention exhibited good in vitro anti-cancer activity towards various human tumor cell lines, according to the results obtained from the 60 human tumor cell line assay performed at National Cancer Institute (NCI), Bethesda, Maryland, U.S.A
Table 1 presents in vitro cell line activity expressed as IC50 values for various 20(S)-camptothecin C-ring analogues prepared in the present invention.
Charts 1 to 3 present the data compiled based on NCI
mean graphs for total growth inhibition (TGI) of different types of human cancer cell lines for the compounds prepared in the examples 27, 28 and 43.
Similar data compiled for topotecan based on NCI mean graph is also included for comparison purposes. The data presented in Tables 2 and 3 shows that the C-ring analogues of 20(S)-camptothecin prepared in the present invention exhibited anti-tumor activity equal or superior to topotecan towards certain cell lines of different cancer cell panels. Table 4 presents the data obtained for the~compound prepared in the example 32 against AIDS related lymphoma(ARL) cell lines. All the compounds used in the NCI's in vitro anti-cancer screening programme are mixtures substantially containing both the diastereomers having 20(S),5(S) and 20(S),5(R) configurations in varied ratios.
The results shown in charts 1 to 3 and tables 1 to 4 were obtained from conducting experiments according to U.S. National Cancer Institute (NCI) protocols as given below:
Each test compound was screened against a battery of 60 human cell lines obtained from eight organs. In a typical procedure, the cell suspensions that were diluted according to the particular cell type and the expected target cell density (5000-40,000 cells per well based on cell growth characteristics) were added into 96-well microtiter plates. Inoculates were allowed a preincubation period of 24h at 37°C. for stabilization. Dilutions at twice the intended test concentrations were added at time zero in 100-fr.l aliquots to microtiter plate wells. Usually test compounds were evaluated at five 10-fold dilutions.
The highest well concentration used in the test is 10-4M. The cells are then incubated in the presence of drug.(the test compound) for further 48h in 5~ C02 atmosphere and 100 humidity. At the end of this time, the adherent cells are fixed to the plate by means of trichloroacetic acid, and after a number of washes, the cell layer is treated with the protein stain Sulforhodamine B. The optical density which is proportional to protein mass, is then read by automated spectrophotometric plate readers at a wavelength of 515 nm. Readings are transferred to a microcomputer and final reports are generated using especially developed software.
The compounds of formula 1 of the present invention, and the pharmaceutically acceptable salts thereof as described above, and the compositions containing them, are useful as anti-cancer and anti-viral agents. Administration of the novel active compounds of the formula l, in pure form or in an appropriate pharmaceutical composition can be carried out via any of the accepted modes of administration for serving similar utilities. Thus, administration can be, for example, orally, nasally, parenterally or topically, in the form of solid, semi-solid, lyophilized powder, or liquid dosage forms, such as for example, tablets, suppositories, pills, capsules, powders, solutions, suspensions, emulsions, creams, lotions, aerosols, ointments, injections or the lilte, preferably, in unit dosage forms suitable, for simple ' administration of precise dosages. The compositions will include a conventional pharmaceutical carrier, diluent or excipient and an active novel compound of WO 97!46563 PCT/US97/06962 formula 1 and, in addition, may include either medicinal agents, pharmaceutical agents, carriers, adjuvants, etc.
The invention is described in detail with specific examples given below, which are provided by way of illustration only and should not be considered to limit the scope of the invention.
TABLE 1:
l0 S . NO. COMPOUND IC5 0 ( fcm) a 1. 5-Methoxycamptothecin* 8.5 2. 5-Ethoxycamptothecin* 9.54 3. 5-n-Butoxycamptothecin* 6.16 4. 5-(2'-Hydroxyethoxy)camptothecin 1.51 5. 5-(2'-Chloroethoxy)camptothecin 4.57 6. 7-Ethyl-5-ethoxycamptothecin* 1.41 7. 9-Methoxy-7-ethyl-5-ethoxycamptothecin 2.13 8. 7-Ethyl-5-chloroethoxy camptothecin 2.75 9. 7-Ethyl-5-aminoethoxy camptothecin 18.6 10. 7-Ethyl-5-pyrollidinoethoxy camptothecin 18.6 11. 7-Ethyl-5-piperidinoethoxy camptothecin >30 12. 7-Ethyl-5-N,N-dimethylaminoethoxy camptothecin 13.8 13. 7-Ethyl-5-N,N-dimethylaminopropoxy camptothecin >30 14. 9-Methoxy-5-ethoxy camptothecin 2.45 15. 5-Trifluoroethoxy camptothecin 1.82 16. 5-Aminoethoxy camptothecin 30.0 17. 7-Ethyl-5-trifluoroethoxycamptothecin 7.41 18. 7-Ethyl-5-(2'-hydroxyethoxy)camptothecin 4.78 19. 5-Fluoroethoxycamptothecin 1.58 20. 10-Hydroxy-5-trifluoroethoxy camptothecin 0.38 21. 9-Nitro-5-trifluoroethoxy camptothecin 0.46 22. 10-Hydroxy-5-(2'hydroxyethoxy)camptothecin 8.12 23. 9-Nitro-5-(2'-hydroxyethoxy)camptothecin 7.94 24. 7-Ethyl-5-fluoroethoxy camptothecin 4.36 WO 97!46563 ~'CT/US97/06962 25. 5-Methoxyethoxy camptothecin 2.23 26. 9-Nitro-5-methoxyethoxy camptothecin 2.04 27. 12-Nitro-5-ethoxy camptothecin >30 28. 12-Nitro-5-hydroxy camptothecin >30 29. 9-Amino-5-methoxy camptothecin 6.76 30. 9-Hydrory-5-ethoxy camptothecin 6.68 a 1050 = the mean value ofthe minimum drug concentration (hum) of the agent required to produce 50~
l0 cell growth inhibition (GI50) against NC2's 60 human tumor cell line assay.
'* represents C-5 substituted camptothecin derivatives known in the literature.

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CELL NAME GI50* TG2**

CCRF - CEM 0.318 1.83 RL 0.463 3.94 .

488 0.246 2.28 AS 283 0.268 0.678 PA 682 0.456 7.23 SU-DHL-7 0.609 3.51 * GI50 refers to the minimum concentration(~,m) of the text compound required for 50~ cell Growth Inhibition.
**TGI refer to minimum concentration (hum) of the text compound required for the Total Growth Inhibition.

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Preparation of 5-methoxyeamptothecin (known compound) Step 1: To a mixture of 20(S)-Camptothecin of the formula 3 (2g), ferric chloride (2g), dissolved in 80mI
of methanol, lOml of sulfuric acid was added dropwise and continued heating at 70°C. for 24h. Excess acid and methanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 1.8g of yellowish powder containing 5-methoxycamptothecin and 5-hydroxy camptothecin in the ratio of 5:1.
Step 2: Separation of the mixture by silica gel column chromatography using methanolchloroform solvent mixture as eluent afforded 1.5g of 5-methoxycamptothecin and 300mg of 5-hydroxycamptothecin. Analytical data for 5-methoxycamptothecin: mp: 156°C.; [x~D at 28°C. -+ 41.?4 (c 0.103, CHC13); IR: 3426, 1747, 1664, 1616, 1228, 1155, 1046, 762 cm-1; 1H NMR (CDCI3, 200MHz): 8 8.42(s, 1H), 8.26(d, J=8Hz, 1H), 7.96(d, J=8Hz, IH), 7.88 (t, J= 6.8Hz, 1H), 7.68(t, J=6.8Hz, 1H), 7.58(s, 0.5H), 7.54(s, 0.5H), 6.95(s, 0.5H), 6.80(x, 0.5H), 5.74(d, J=16.5Hz, 0.5H), 5.72 (d, J=16.5Hz, 0.5H), 5.25(d, J=16.5Hz, 1H), 3.75(s, 1H), 3.70(s, 1.5H), 3.50(s, 1.5H), 2.01-1.82(m, 2H), 1.06(t, J=7Hz, 3H);
Mass (m/z): 379(M+H), 348, 319.

Preparation of 5-Hydroxycamptothecin (known compound) Step 3: Preparation of 5-methox~ camptothecin: 5-Methoxycamptothecin of the formula 2 where R1=R2=R3=R4=R5=H, R6=Me was prepared from 20(S)-camptothecin of the formula 3 as described in the example 1.
Step 2: 1.5g of 5-methoxycamptothecin of the formula 1 WO 97/46563 PCT/gJ597/06962 where R1=R2=R3=R4=R5=H, R6=Me was dissolved in 50m1 of methanol and treated with 50 ml of 50~ HC1. The solution was heated to reflux for 30h. At the end, excess water and methanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 1.2g of 5-hydroxycamptothecin after purification over silica gel column chromatography using a solvent mixture of ethyl acetate and chloform;
mp: 220°C.; [oc]D at 26°C. _ +28.00 (c 0.1 in CHC13);
IR: 3367, 1749. 1658, 1591, 1159, 1046 cm-1; 2H NMR
(CDC13+DMSO-d6, 200MHz): S 8.50 (s, 1H), 8.20 (d, J=
8Hz, 1H}, 7.94(d, J=8Hz, 1H), 7.85(t, J=6.8Hz, 1H), 7.64(t, J=6.8Hz, 1H), 7.58 (s, 0.5H), 7.56(s, 0.5H), 7.06 (s, 0.5H} , 7.01 (s, 0.5H) , 6.95 (br d, 1H, D20 exchangeable), 5.67(d, J= 16.5Hz, 1H), 5.25(d, J=16.5Hz, 1H), 5.05 (br d, 1H, D20) exchangeable), 2.05-1.86 (m, 2H) , 1. 06 (t. J=7Hz, 3H) ; Mass (m/z) 364(M+1), 348, 320, 277, 236, 91, 57.

Preparation of 5-Ethox~-7-ethZrlcamt~tothecin (known compound}
Step 1: To a mixture of 7-ethyicamptothecin of the formula 2 where R1=R2=R3=R4=H, R5=Et(1.5g}, ferric chloride (1.35g), dissolved in 150m1 of ethanol, 9ml of sulfuric acid was added dropwise and continued heating at 85°C. for 30h. Excess acid and ethanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 1.6g of brownish powder containing 5-ethoxy-7-ethylcamptothecin and 5-hydroxy-7-ethylcamptothecin in the ratio of 10:1. ' Step 2: Separation of the mixture by column chromatography gave lg of 5-ethoxy-7-ethyl-camptothecin and 100mg of 5-hydroxy-7-ethylcamptothecin; mp: 150°C;

WO 97146563 PCT/US97l06962 [oc]D at 27°C. - + 10.526 (c 0.085, CHC13); IR: 3419, 1751, 1662, 1613, 1157, 1075, 1050, 764 em-l~ 1H NMR
. (CDC13, 200MHz) : S 8.20 (d, J=8Hz, 1H) , 8.15 (d, J=8Hz, 1H}, 7.81(t, 3=6.8Hz, 1H), 7.66(t, 7.3Hz, 1H), 7.54(s, 0.5H), 7.51(s, 0.5H), 7.01(s, 0.5H), 6.89(s, 0.5H), 5.72(d, J=16.5Hz, 0.5H), 5.71(d, J=16.5Hz,0. 5H), 5.28(d, J=16.5Hz, 0.5H), 5.26(d, J=16.5Hz, 0.5H), 4.3-3.6(m, 3H), 3.5-3.1 (m, 2H), 2.05-1.71(m, 2H), 1.45(t, J=7.5Hz, 3H), 1.06(t, J=7Hz, 3H).

Preparation of 5-Hydroxv-7-ethylcam~tothecin (known compound) Step 1: Preparation of 5-Ethoxy-7-ethvlcamptothecin:
5-Ethoxy-7-ethyl.camptothecin of the formula 1 where R1=R2=R3=R4=H, R5=R6=Et, was prepared from 20(S)-camptothecin of the formula 2 as described in the example 3.
Step 2: 50 ml of 25~ H2S04 was added to l.Og of 5-ethoxy-7-ethylcamptothecin of the formula 1 where R1=R2=R3=R4=H, R~=R6=Et, dissolved in 30m1 of ethanol and heated to reflex for 30h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 700mg of 5-hydroxy-7-ethylcamptothecin after purification over silica gel column chromatography mp: 252°C.; IR:
3349, 1752, 1656, 1605, 1159, 1054, 766 em-l; Partial data of 1H NMR (CDC13 + DMSO-d6) 8 7.19(br s, 1H, D20 exchangeable), 7.15(s, 0.5H), 7.05(s, 0.5H), 5.75(br s, 1H, D20 exchangeable), 5.65(d, J=16.5Hz, 1H), 5.25 (d, J=16.6Hz, 1H), 3.52-3.19(m, 2H), 1.45(t, J= 7.5Hz, 3H), 1 . 02 (m, 3H) .

Preparation of 5-Ethoxy-9-methoxycamptothecin of the formula 1 where RI=OMe, R2=R3=R4=R5=H, R6=Et Step 1: To a mixture of 9-methoxycamptothecin of the formula 2 where R~=OMe, R2=R3=R4=R6=H(lg), ferric chloride (500mg), dissolved in 50m1 of ethanol l0ml of sulfuric acid was added dropwise and continued heating at 85°C. for 22h. Excess acid and ethanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded dark brownish powder.
Step 2: Purification of the above residue over column chromatography using silica gel furnished 500mg of 5-ethoxy-9-methoxycamptothecin of the formula 1 and 300mg of 5-hydroxy-9-methoxycamptothecin; mp: 235°C.; [«]D at 30°C.= + 34.1$ (c 0.093, MeOH); IR: 3436, 748, 1665, 1619, 1461, 1366, 1093, 814cm-1; 1H NMR (CDC13, 200MHz):
8 8. 81 (s, 1H) , 7 .78 (m, 2H) , 7 .53 (d, J=5 . 5Hz, 1H) , 6.96 (d, J=7Hz, 1H) , 6.88 (s, 1H) , 6.77 (s, 1H) , 5.72 (d, J=l6Hz, 0.5H), 5.75(d, J=l6Hz, 0.5H), 5.27(d, J=l6Hz, 1H), 4.24-3.90(m, 2H), 4.06(s, 3H), 3.80(s, 1H, D2O
exchangeable), 1.90(m, 2H), 1.31(m,3H), 1.01(m, 3H);
Mass (m/z): 422(M+1), 394, 378, 350, 305, 98, 57.

Preparation of 5-Hy~droxy-9-methox~rcamptothecin (known compound) Step 1: Initially 5-Ethoxy-9-methoxycamptothecin of the formula 1 where R1=OMe, R2=R3=R4=R5=H, R6=Et, was prepared as described in the example 5.
Step 2: 25 ml of 80~ HCl was added to 560mg of 5-ethoxy-9-methoxycamptothecin of the formula 1 where Rl=OMe, R2=R3=R4=R5=H, R6=Et, dissolved in 25m1 of ethanol and heated to reflex for 16h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 520mg of 5-hydroxy-9-methoxy-camptothecin after purification over silica gel column chromatography; mp: 162°C.; [a7D at 30°C. - + 39.68 (c 0.012, MeOH); IR: 3398, 1749, 1656, 1616, 1577, 1465, 1383, 1154, cm-I; ZH NMR {CDC13+ DMSO-d6): b 8.81(s, 1H), 7.81-7.61(m, 2H), 7.50(d, J=5.5Hz, 1H), 7.12-6.71 (m, 2H), 5.70(d, J=l6Hz, 1H}, 5.30(d, J=l6Hz, 1H), 4.06(s, 3H), 1.98-1.75(m,2H), 1.10-0.98(m,3H); Mass (m/z): 394(M+1), 377, 348, 266, 149, 88, 57.

Preparation of 5-Ethoxy-9-methoxy-7-ethylcamptothecin of the formula 1 where R1=OMe,R2=R3=R4=H,RS=R6=Et Step 1: To a mixture of 9-methoxy-7-ethylcamptothecin of the formula 2 where R1=OMe,R2=R3=R4=H, R5=Et{100mg), ferric chloride (100mg), dissolved in 32m1 of ethanol 2m1 of sulfuric acid was added dropwise and continued heating at 85°C. for 4h. Excess acid and ethanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 120mg of residue containing 5-ethoxy-9-methoxy-7-ethylcamptothecin and 5-hydroxy-9-methoxy-7-ethylcamptothecin in the ratio of 1:4.
Step 2: Separation of the mixture by column chromatography using the solvent mixture of ethyl acetate-chloroform gave l5mg of 5-ethoxy-9-methoxy-7-ethylcamptothecin of the formula 1 and 55mg of 5-hydroxy-9-methoxy-7-ethylcamptothecin; mp: 240°C.; 2R:
3443, 1747, 1663, 1609, 1458, 1254, 1160, 1074 cm-1; 1H
NMR (CDC13, 200MHz): 8 7.80-7.68(m, 2H), 7.50(s, 0.5H), 7.47{s, 0.5H), 7.01(s, 0.5H), 6.98(d, J=8Hz, 1H), 6.89(s, 0.5H}, 5.76(d, J=l6Hz, 1H). 5.28(d, J=l6Hz, 0.5H), 5.26(d, J=l6Hz, 0.5H), 4.25-3.8.1(m, 2H), 4.02(s, 3H), 3.72-3.28(m, 2H}. 3.15(br s, 1H, D20 exchangeable), 2.02-1.82(m, 2H), 1.3?-1.33(m, 3H), 1.05-0.95(m, 3H); Mass (m/z): 451(M+1), 406, 377, 362, 347, 331, 261, 181, 149, 97.

Preparation of 5-Hydroxy-9-methoxy-7-ethylcamptothecin of the formula 13 where R1=OMe, R2=R3=R4=H,R5=R6=Et Step 1: Initially 5-Ethoxy-9-methoxy-7-ethylcamptothecin of the formula 1 where R1=OMe, R2=R3=R4=H,RS=R6=Et, was prepared as described in the example 7.
Step 2: 5 ml of 50~ HCl was added to 100mg of 5-ethoxy-9-methoxy-7-ethylcamptothecin of the formula 1 where Rl=OMe, R2=R3=R4=H,R5=R6=Et, dissolved in 5ml of ethanol and heated to reflux for 26h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine anal dried over anh.sodium sulfate. Concentration of the solvent afforded 80mg of 5-hydroxy-9-methoxy-7-ethylcamptothecin of the formula 13 after purification over silica gel column chromatography; mp: 242°C.; IR:
3440, 1742, 1660, 1610, 1456, 1250, 1160 cm-I.

Preparation of 5-Ethoxycamptothecin (kn.own compound) Step 1: To a mixture of 20(S)-Camptothecin of the formula 3 (lg), ferric chloride (lg), dissolved in 50m1 of ethanol l2ml of BF3-etherate was added dropwise and continued heating at 85°C. for 40h. Excess acid and ethanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate.
Concentration of the solvent afforded lg of yellowish powder containing 17-ethoxycamptothecin and 5-hydroxycamptothecin in the ratio of 6:1.
Step 2: Separation of this mixture by silica gel column chromatography using ethyl acetate hexane solveat mixture as eluent afforded 700mg of 5-ethoxycamptothecin and 120mg of previously prepared 5-hydroxycamptothecin; mp: 140°C.; [«]D, at 28°C. -+ 29.703 (c 0.101, CHC13); IR: 3423, 1746, 1663, 1616, 1155, 1070, 1040 cm-1; Partial 1H NMR data in CDC13: b 6.9(s, 0.5H), 6.78(s, 0.5H), 4.25-3.85(m, 2H), 3.70(s, 1H), 2.00-1.80(m, 2H), 1.40-1.22(m, 3H), 1.12-0.98(m, 3H); Mass (m/z): 393 (M+1), 378, 362, 348, 319, 247, 219, 57.

Preuaration of 5-butoxycamptothecin (known compound) Step 1: To a mixture of~20(S)-Camptothecin of the formula 3(500mg), ferric chloride (500mg), dissolved in 15m1 of n-butanol, sulfuric acid was added dropwise and continued heating at 100°C. for 20h. Excess acid and n-butanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate.
Concentration of the solvent afforded powdery material.
Step 2: Purification of the above material by silica gel column chromatography using ethylacetate-hexane solvent mixture as eluent afforded 300mg of 5-butoxycamptothecin and 50mg of previously prepared 5-hydroxycamptothecin; mp: 82°C.; C«]D at 28°C. - +28.00 (c 0.1, CHC13); Partial 1H NMR data in CDC13: b 6.92(s, 0.5H), 6.79(s, 0.5H), 4.12-3.75(m, 2H), 3.80(br s, 1H, D20 exchangeable), 2.00-1.82(m, 2H), 1.75-1.52 (m, 2H), 1.50-1.29(m, 2H), 1.15-0.82(m, 6H); Mass (m/z):
422(M+1), 363, 348, 319, 84, 51.

Preparation of 5-Ethoxy-9-hydroxyeamptothecin of the formula 1 where RI=OH,R2=R3=R4=R5=H,R6=Et Step 1: To a mixture of 9-hydroxycamptothecin of the formula 2 where Rl=OH, R2=R3=R4=R5=H, (200mg), ferric chloride (250mg), dissolved in 40m1 of ethanol 1.5m1 of sulfuric acid was added dropwise and continued heating ' at 85°C for 26h. Excess acid and ethanol were removed under vacuum and the residue was extracted with 5~
methanol-chloroform. Organic layer was washed with water, brine and dried over anh.sodium sulfate.
Concentration of the solvent afforded 170mg of residue containing 5-ethoxy-9-hydroxycamptothecin and 5,9-WO 97/46563 PCT/iJS97/06962 dihydroxycamptothecin in the ratio of 3:1.
Step 2: Separation of the mixture by column chromatography using the solvent mixture of ethyl acetate-chloroform gave 75mg of 5-ethoxy-9-hydroxycamptothecin of the formula 1 along with 25mg of 9,5-dihydroxycamptothecin; mp: 230°C.; IR: 3400, 2920, 1745, 1663, 1597, 1360, 1280, 1228, 1157, 1083, 902, 816 cm-1; 1H NMR (CDC13): b 8.83(s,lH), 7.78 {d, J=6.8Hz, 1H), 7.67-7.56 (m, 2H), 7.01(s, 0.5H), 6.98(s, 0.5H), 6.91(s, 0.5H), 6.81(s, 0.5H), 5.70(d, J=l6Hz, 1H), 5.33(d, J~36Hz, 1H), 4.15-3.91(m, 2H), 1.90(m, 2H), 1.05(t, J=7Hz, 3H); Mass (m/z): 409(M+1), 364, 335, 320, 291, 267, 263, 221, 206, 171, 159, 129, 111, 98. 85.

Preparation of 9 5-Dihydroxycamptothecin of the formula 13 where RI=OH,R2=R3=R4=R5=H
Step l: Initially 5-Ethoxy-9-hydroxycamptothecin of the formula 1 where R1=OH, R2=R3=R4=R5=H, R6=Et, was prepared as described in the example 11.
Step 2: 25 ml of 80$ HCl was added to 560mg of 5-ethoxy-9-hydroxy camptothecin of the formula 1 where R1=OH, RZ=R3=R4=R5=H, R~=Et, dissolved in 25m1 of ethanol and heated to reflex for Z6h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 320mg of 9,5-dihydroxycamptothecin of the formula 13 after purification over silica gel column chromatography; mp: 102°C.; IR: 3400, 1744, 1659, 1594r 1462, 1361, 1280, 1229, 1049, 820 cm-I': ZH ' NMR (DMSO-d6): S 10.82 (s, 1H, D20 exchangeable), 7.63-7.69(m, 2H), 7.22(s, 0.5H), ?.19(s, 0.5H), 7.11(d, J=7Hz, 1H), 6.98(s, 0.5H), 6.95(x, 0.5H), 6.50(s, 1H, D20 exchangeable), 5.42(s, 2H), 1.89(m, 2H), 0.90(t, J=7Hz, 3H); Mass(m/z): 380(M+1), 320, 305, 293, 264, Preparation of 5-Ethoxv-9-h~sdroxv-7-ethylcamp tothecin of the formula 1 where RZ=OH. R2=R3=R4=H. R5=R6=Et Step l: To a mixture of 9-hydroxy-7-ethylcamptothecin of the formula 2 where Rl=OH, R2=R3=R4=H R5=Et,(150mg), ferric chloride (150mg), dissolved in 30m1 of ethanol, 2m1 of sulfuric acid was added dropwise and continued heating at 85°C. for 40h. Excess acid and ethanol were removed under vaccum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 120mg of residue containing 5-ethoxy-9-hydroxy-7-ethylcamptothecin and 9,5-dihydroxy-7-ethylcamptothecin in the ratio of 5:1.
Step 2: Separation of the mixture by column chromatography using the solvent mixture of acetone-chloroform gave 85mg of 5-ethoxy-9-hydroxy-7-ethylcamptothecin of the formula 1 and l5mg of 5,9-dihydroxy-7-ethylcamptothecin; mp: 240°C.; IR; 3500, 2976, 1749, 1662, 1588, 1555, 1461, 1390, 1147, 1079, 921 cm-Z; 1H NMR (DMSO-d6): b 10.77(s, 1H D20 exchangeable), 7.62-7.67(m,2H), 7.57(d, J=8Hz, 1H), 7.08-7.18(m,2H), 6.50(s, 1H, D20 exchangeable), 5.39(s, 2H), 4.08(m, 2H), 3.42(m, 2H), 1.87(m, 2H), 1.35(t, J=7Hz, 3H); 0.87(t, J=7Hz, 3H); Mass (m/z) . 437(M+1), 392, 363, 348, 333, 291, 261, 246, 219, 191, 149, 119, 89.

Preparation of 9,5-Dihydroxy-7-ethylcamptothecin of the formula 13 where R1=OH. R2=R3=R4=H, R~=Et Step 1: Initially 5-Ethoxy-9-hydroxy-7-ethylcamptothecin of the formula 1 where R~=OH, R2=R3=R4=H, R5=R6=Et, was prepared as described in the example 13.
Step 2: 35 ml of 80~ HC1 was added to 560mg of 5-ethoxy-9-hydroxy-7-ethylcamptothecin of the formula 1 where RZ=OH, R2=R3=R4=H, R5=R6=Et, dissolved in 25m1 of ethanol and heated to reflex for 16h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 380mg of 5,9-dihydroxy-7-ethylcamptothecin of the formula 13 of ter purification over silica gel column chromatography; mp: I65°C.. IR
3351, 2929, 1744, 1657, 1606, 1460, 1218, 1162, 1035, 872 cm 1: 1H NMR (DMSO-d6): b 10.62 {s, 1H, D20 exchangeble), 7.60-7.57(m, 2H) 7.16-7.00(m, 3H), 5.40(x, 2H, 3.42(q, J=7.6Hz, 2H), 2.08(m,2H), 1.33(t, J=7Hz, 3H), 0.89(t, J=7Hz, 3H); Mass(m/z): 408(M+1), 380, 336, 319, 291, 267, 235, 219, 185, 127, 99, 83.

Prer~aration of 9-vitro-5-ethoxy camptothecin of the formula 1 where Rl=N02. R2=R3=R4-R5=H, R6=Et Step 1: To a mixture of 9-nitrocamptothecin of the formula 2 where R1=N02 R2=R3=R4=R5=H, (lg), ferric chloride (lg), dissolved in 100m1 of ethanol, 10m1 of sulfuric acid was added dropwise and continued heating at 85°C. for 24h. Excess acid and ethanol were removed under vaccum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 900mg of yellowish powder.
Step 2: Purification of the above solid material over silica gel column chromatography using acetone-chloroform solvent mixture as eluent furnished 700mg of 9-vitro-5-ethoxycamptothecin of the formula 1 and 80mg of 9-vitro-5-hydroxycamptothecin of the formula 13 where R1=N02, R2=R3=R4=R5=H; mp: 202°C.; IR (KBr): 3474, 1743, 1668, 1622, 1526, 1344, 1154, 1073, 831cm-1; 1H
NMR(CDC13, 200 MHz): 8 9.23 (s, 1H), 8.52(d, J=9Hz, 1H), 8.47(d, J=9Hz, IH), 7.92{t, J=8.2Hz, 1H), 7.55(s, 1H) , 6.91 (s, IH) , 5.7I (d, J=I6Hz, IH) , 5.28 (d, J=l6Hz, 1H), 4.39-3.98(m, 2H), 3.75(br s, 1H, D20 exchangeable), 3.99-1.79(m, 2H), 1.32(t, J=7Hz, 3H), 1.04(t, J=7Hz, 3H); Mass{m/z): 438(M+1), 407, 393, 364, 349, 319, 262, II8.

Preparation of 12-Nitro-5-ethoxycam~tothecin of the formula 1 where IO R1=RZ=R3=R5=H, R4=N02, R6=Et Step 1: To a mixture of 12-nitrocamptothecin of the formula 2 where R4=N02 R1=R2=R3=R5=H, (2g), ferric chloride (2g), dissolved in 150m1 of ethanol, l5ml of sulfuric acid was added dropwise and continued heating at 85°C. for 24h. Excess acid and ethanol were removed under vaccum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent gave a gummy solid material.
Step 2: Purification of the above residue over silica gel column chromatography using acetone-chloroform solvent mixture as eluent afforded 1.4g of yellowish powder containing 12-vitro-5-ethoxycamptothecin of the formula 1 and 100mg of I2-vitro-5-hydroxycamptothecin of the formula 13 where R4=N02, R~=R2=R3=R5=H; mp:
250°C.; IR (KBr): 3450, 1750, 1666, 1618, 1525, 1357, 1154, 1042, 766 cm-1; ~H NMR (CDC13, 200 MHz); 8 8.49(s, 1H), 8.17(d, J=9Hz, 2H), 8.I4(d, J=9Hz, 1H), 7.75(t, J=8.2Hz, 1H), 7.54( s, 1H), 6.95(s, 0. 5H), 6.82(s, 0.5H), 5.7I(d, J=l6Hz, 1H), 5.26(d, J=I6Hz, 1H), 4.31-3.91(m, 2H), 3.75(m, br s, 1H, D20 exchangeable), 2.05-1.81(m, 2H), I.35(I, J=7Hz, 3H), 1.05(1, J=7Hz, 3H};
Mass {m/z) . 438(M+1), 420, 393, 376, 364, 349, 3I9, 84.
3 5 EXAMPLE 7.7 Preparation of 10-hydroxY-5-ethoxy camptothecin (Itnown compound) WO 97!46563 PCT/LTS97/06962 Step 1: To a mixture of 10-hydroxycamptothecin of the formula 2 where R2=OH, R1=R3=R4=R5=H, (200mg), ferric chloride (200mg), dissolved in l0ml of ethanol, l.5ml of sulfuric acid was added dropwise and continued heating at 85°C. for 24h. Excess acid and ethanol were removed under vaccum and the residue was extracted with 5~ methanolethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate.
Concentration of the solvent afforded yellow solid material.
Step 2: Purification of the above solid over silica gel column chromatography using acetone-chloroform solvent mixture as eluent provided 100mg of 20-hydroxy-5-ethoxycamptothecin of the formula 1 and 20mg of 10.5-dihydroxycamptothecin of the formula 13 where R2=OH, R1=R3=R4=R5=H; mp; 165°C.; IR (KBr) ; 3384, 1747, 1662, 1608, 1229, 1044, 831 cm-1; 1H NMR(CDC13+ DMSO, 200MHz):
9 . 8 (1H, br s, D20 exchangeable) . 8 .25 (s, 1H) , 8.05 (d, J=6Hz, 1H), 7.56-7.39(m. 2H), 7.25(s, 1H), 6.85(s, 0.5H), 6.70(s, 0.5H), 5.58(d, J=l6Hz, 1H), 5.35 (d, J=l6Hz, 0.5H), 5.21(d, J=l6Hz, 0.5H), 4.35-3.75(m, 4H), 3.50( br s, 1H, D20 exchangeable), 2.10-3.78 (m, 2H), 1.22 (t, J=7Hz, 3H), 1.05(t, J=7Hz, 3H); Mass (m/z):
409(Mfl), 392, 364, 349, 335, 320, 291, 235, 117, 84.

Preparation of 10-HSrdroxy-7-et~l-5-ethox~r camntothecin (Icnown compound) Step 1: To a mixture of 10-hydroxy-7-ethylcamptothecin of the formula 2 where R2=OH, R1=R3=R4=H, R5=Et, (200mg), ferric chloride (200mg), dissolved in lOml of ethanol, 1_7ml of sulfuric acid was added dropwise and continued beating at 80°C. for 20h. Excess acid and ethanol were removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate.
Evaporation of the solvent gave a solid material.

WO 97/46563 PCTlUS97/06962 Step 2: Purification of the above solid residue over silica gel column chromatography using acetone-chloroform solvent mixture as eluent afforded 85mg of 10-hydroxy-7-ethyl-5-ethoxy-camptothecin as yellowish powder and 20mg of 10,5-dihydroxy-7-ethylcamptothecin of the formula 13 where R2=OH, Rl=R3=R4=H, R6=Et; mp:
190°C.; TR (ICBr) : 3277, 1.746, 1660, 1599, 1231, 1078, 800 cm-1; 1H NMR (CDC13+ DMSO): s 9.6(br s, 1H, D20 exchangeable), 8.01{ d, J=8.7Hz, 1H}, 7.51-7.35(m, 3H), 6.92(s. 0.5H}. 6.80(s, 0.5H), 5.66(d, J=l6Hz, 1H), 5.22(d, J=l6Hz, 1H), 3.85-3.65(m, 2H), 3.35-2.95(m, 2H), 1.95-1.75(m, 2H), 1.37(t, J=7.4Hz, 3H), 1.17(t, J=7.2Hz, 3H), 0.99(t, J=7.4Hz, 3H); Mass (m/z}:
437(M=1), 392, 363, 348, 333, 291, 147, 84.

Preparation of 9-amino-5-ethoxycamptothecin of the formula 1 where RZ=NH2.R2=R3=R4=R5=H, R6=Et Step 1: To a mixture of 9-aminocamptothecin of the formula 2 where R1=NH2 R2=R3=R4=R5=H, (120mg), ferric chloride (112mg), dissolved in l0ml of ethanol, 1.5m1 of ethereal borontrifluoride(BF3-Et20) was added dropwise and continued heating at 80°C. for 16h.
Ethanol was removed under vaccum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate which upon evaporation of the solvent gave a thick gummy solid.
Step 2: Purification of the above residue over silica gel column chromatography using acetone-chloroform solvent mixture as eluent afforded 65mg of 9-amino-5-ethoxycamptothecin of the formula l; mp: 170°C.; =R
3221, 1744, 1663., 1231, 1157, 1074, 815 em-Z: 1H NMR
(CDC13+DMSO-d6): b 8.69(s, 1H), 7.64{s. 1H), 7..63-7.51{m,2H}. 7.06(d, J=5.41Hz, 1H), 6.90(s, 0.5H), 6.80(s, 0.5H}, 5.65{d, J=l6Hz, 1H), 5.26(d, J=l6Hz, 1H), 4.19-3.98(m, 1H), 3.97-3.78(m, 1H), 2.98(br s, 3H, WO 97/46563 PCTlUS97/06962 D20 exchangeable), 1.95-1.80{m, 2H}, 1.39-1.19(m, 3H), 1.11-0.95 (m, 3H) ; Mass (m/z) : 407 (M+2) , 389, 363, 334, 319, 290, 262, 233 ,101.

Preparation of 9-amino-5-methoxsrcamptothecin of the formula 1 where RZ=NH2. R2=R3=R4=R5=H, R6=Me Step 1: To a mixture of 9-aminocamptothecin of the formula 2 where R1=R2=R3=R4=R5=H, (180mg), ferric chloride (162mg), dissolved in 15m1 of methanol, 2ml of ethereal borontrifluoride(BF3-Et20) was added dropwise and continued heating at 80°C. for 16h. Methanol was removed under vacuum and the residue was extracted with ethylacetate. Organic layer was washed with water, brine and dried over anh.sodium sulfate which upon evaporation of the solvent gave a thick gummy solid.
Step 2: Purification of the above residue over silica gel column chromatography using acetone-chloroform solvent mixture as eluent afforded 125mg of 9-amino-5-methoxycamptothecin of the formula 1; mp: 200°C.; IR:
3364, 2925, 1744, 1660, 1610, 1156, 1081, 813 cm-1; 1H
NMR (CDC13-t- DMSO-d6} : S 8. 82 (s, 1H) , 7. 60 (s, 1H) , 7.63-7.46(m, 2H), 6.97(d, J=7Hz, 1H}, 6.89(s, 0.5H, 6.80(s, 0.5H), 5.6(d, J=l6Hz, 1H), 5.25(d, J=l6Hz, 1H), 3 .57 (s, 1.5H) , 3.46 (s, 1.5H) , 3 .41 (br s, 1H, D20 exchangeable), 3.15(br s, 2H, D20 exchangeable), 2.05-1.89 (m,2H) , 1.01 (t, J=7Hz, 3H) ; Mass (m/z) : 393, (Mtl) , 376, 363, 349, 334, 319, 290, 262, 233, 205, 116.

Prer~aration of 9-Nitro-5-hydroxy camntothecin of the formula 13 where R1=N02. R2=R3=Rg=R5=H
Step 1: Initially 9-vitro-5-ethoxycamptothecin of the formula 1 where R1=N02, R2=R3=R4=R5=H, R6=Et was prepared as described in the example l5.
Step 2: 80 m1 of 50~ HCl was added to l.Og of 9-nitro-5-ethoxycampothecin of the formula 1 where R1=N02, R2=R3=R4=R5=H, R6=Et, dissolved in 20m1 of ethanol and heated to reflex for 30h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 700mg of 9-vitro-5-hydroxycamptothecin of the formula 13 after purification over silica gel column chromatography.; mp: 278°C.; IR (KBr} . 3402, 1744, 1657, 1602, 1533, 1155, 1051, 833 cm-1; 1H NMR (CDC13 +DMSO, 200MFiz) : b 9.28 (s, 1H) , 8.50 (d, J=8. 6Hz, 1H) , 8.45(d, J=8.6Hz, 1H), 7.96(t, J=8.2Hz, 1H), 7.59(s, 0.5H), 7.58(s, 0.5H), 7.12(s, 0.5H), 7.08(s, 0.5H), 5. 67 (d, J=l6Hz, IH) , 5.27 (d, J=l6Hz, 1H) , 1.92 (q, J=7.2Hz, 2H), 1.07(t, J=7Hz, 3H).

Preparation of 10,5-DihYdroxy cam~tothecin (known compound) Step 1: 2nitially 10-Hydroxy-5-ethoxycamptothecin of the formula 1 where R2=OH, R2=R3=R4=R~=H, R6=Et was prepared as described a.n the example 17.
Step 2: 10 ml of 50% HCl was added to 250mg of 10-hydroxy-5-ethoxycamptothecin of the formula 1 where R2=OH, Rl=R3=R4=R5=H, R6-Et, dissolved in l0ml of ethanol and heated to reflex for 20h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 210mg of 10,5-dihydroxycamptothecin of the formula 13 after purification over silica gel column chromatography.; mp: 240°C.; IR (KBr): 3226, 1743, 1659, 1596, 1382, 1231, 1048, 832 cm-1; 1H NMR
(CDC13+DMSO): b 10.0 (br s, 1H, D20 exchangeable), 8.31(s, 0.5H), 8.29(s, 0.5H), 8.05(d, J=6Hz, 0.5H), 7.95(d, J=6Hz, 0.5H), 7.95 (d, J=6Hz, 0.5H), 7.50-7:31(m, 2H), 7.21(x, 1H), 6.95(s, 0.5H), 6.85(s, 0.5H), WO 97/46563 PCTlLTS97/06962 .55 (d, J=l6Hz, 1H) , 5.25 (d, J=l6Hz, 1H) . 3 . 99 (br s, 1H, DZO exchangeable), 2.05-1.81(m, 2H), 1.0(t, J=7Hz, 3H); Mass (m/z): 381(M+1), 352, 336, 320, 264, 149, 83.

5 Preparation of 12-Nitro-5-hvdroxy camptothecin of the formula 13 where R1=R2=R3=R5=H~ R4=N02 Step 1: Initially 12-vitro-5-ethoxycamptothecin of the formula 1 where R4=N02, R1=R2=R3=R5=H, R6=Et was prepared as described in the example 16.
Step 2: 125 ml of 50~ HCl was added to 2g of 12-nitro-5-ethoxycamptothecin of the formula 1 where R4-N02, RI=R2=R3=R5=H, R6=Et, dissolved a.n 30m1 of ethanol and heated to reflux for 24h. At the end, excess water and ethanol were removed as an azeotropic mixture and the residue was extracted with ethylacetate. Organic layer was washed with brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 1.5g of 12-vitro-5-hydroxycamptothecin of the formula 13 after purification over silica gel column chromatography; mp:
247°C.: =R (KBr): 3371, 1746, 1664, 1602, 1532, 1380, 1048, 829 cm-1: 1H NMR (CDC13, 200MHz) . b 8.58(s, 1H), 8.17 (d, J=9Hz, 1H) , 8.12 (d, J=9Hz, 1H) , 7 .74 (t, J=8.2Hz, 1H), 7.58(s, 1H), 7.12(s, 0.5H), 7.08(s, 0.5H), 5.71(d, J=l6Hz, 1H), 5.26(d, J=l6Hz, 1H), 3.90(br s, 1H, D20 exchangeable), 1.99-1.85(m, 2H), 1.05(t, J=7Hz, 3H); Mass (m/z) . 409(M+1), 393, 380, 363, 348, 333, 318, 149, 85.

Preparation of 5-(2'-Chloroethoxy)camptothecin of the formula 1 where Rl=R2=R3=R4=R5=H, R6=CHZCHZCl Step 1: To a mixture of 20(S)-Camptothecin of the formula 3 (lg) , ferric chloride (lg) , dissolved in 25m1 of 2-chloroethanol 5m1 of sulfuric acid was added dropwise and continued heating at 90°C. for 24h.
Excess acid and 2-chloroethanol were removed under WO 97146563 PCTlCTS97/06962 vacuum and the residue was extracted with ethylacetate.
Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 1.2g of brownish solid Step 2: The above solid was purified by column chromatography to afford 650mg of 5-(2'-chloroethoxy)camptothecin of the formula 1 and 150mg of previously prepared 5-hydroxy-camptothecin of the formula 13 where R1=R2=R3=R4=R5=H; mp :202°C.; [a)D at 28°C.=+5.37 (c 0.093, CHC13); TR: 3354, 1744, 1662, 1622, 1223, 1160, 1090, 1044, 752, 663 cm-2; Partial 1H
NMR data in CDC13: b 6.92 (s. 0.5H), 6.82(s, 0.5H), 4.51(ta J=SHz, 1.5H), 4_38(t, J=5Hz, 1_5H), 3.75(s, 1H, D20 exchangeable), 3.85-3.58(m, 2H), 2.00-1.78(m, 2H), 1.06(to J=7.5Hz, 3H); Mass(m/z): 426(M+1), 391, 377, 363, 348, 319, 105, 84, 51.

Preparation of 5-trifluoroethoxycamptothecin of the formula 1 where Rl=Ra=R3=R4=R5=H. R6=CH2CF3 Step 1: To a mixture of 20(S)-Camptothecin of the formula 3 ( 0 . 5g) , f erric chloride ( 0 . 5g) , dissolved a.n 18m1 of 2,2,2-trifluoroethanol, sulfuric acid was added dropwise and continued heating at 80°C. for 24h.
Excess acid and trifluoroethanol were removed under vacuum and the residue was extracted with ethylacetate.
Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent afforded 600mg of solid material.
Step 2: The above solid material was purifsed by column chromatography to give 250mg of 5-trifluoroethoxycamptothecin of the formula 1 along with 150mg of previously prepared 5-hydroxycamptothecin of the formula 13; mp 188°C. ; IR: 3438, 1748, 1667, 1620, 1160, 1106, 1003 cm-1; Partial ZH NMR data in CDC13: 8 6.84 (s, 0.5H), 6.75(s, 0.5H), 5.21-4.90(m, 1H), 4.60-4.38(m,2H), 3.70(s, 1H, D20 exchangeable), 2.0-1.79(m, 2H), 1.15-0.99(m, 3H); Mass (m/z):447 (M+1), 378, 348, 304, 111, 69.

Preparation of 5-(2'-Hvdroxyethoxy)camntothecin of the formula 1 where Rl=R2=R3=R4=R5=H, R6=CH2CH20H
Step 1: To a mixture of 20(S)-Camptothecin of the formula 3 (lg), ferric chloride (1g), dissolved in lOml of ethylene glycol, 5ml of sulfuric acid was added dropwise and continued heating at 70°C. for 36h.
Excess acid and ethylene glycol were removed under vacuum and the residue was extracted with ethylacetate.
Organic layer was washed with water, brine and dried over anh.sodium sulfate. Concentration of the solvent gave l.lg of yellowish powder.
Step 2: The solid obtained as above was subjected to column purification using ethylacetate-hexane solvent mixture to afford 700mg of 5-(2'-Hydroxyethoxy)campto-thecin of the formula 1 and 200mg of previously prepared 5-hydroxycamptothecin of the formula 13 where R1=R2=R3~Rg=R5=H; mp: 190°C; [a]D at 26°C. _ + 28.30 (c 0.106, CHC13); IR: 3300, 3285, 1745, 1665, 1620, 1605, 1227, 1160, 1112, 1047 cm-l; Partial ZH NMR data in CDC13: b 7.01 (s, 0.5H), 6.92 (s, 0.5H), 4.30-3.71(m, 4H), 3.75( br s, 2H, D20 exchangeable), 2.0-1.79(m 2H), 1.15-0.95 (m, 3H); Mass (m/z): 408(M+1), 390, 378, 364, 348, 319, 101, 76.

Preparation of 10-Hydroxy-5-trifluoroethoxycamptothecin of the formula 1 where Rl=R3-R4=R5=H, R2-OH, R6=CH2CF3 Step 1: Initially 10,5-dihydroxycamptothecin of the formula 13 where RZ=R3=R4=R5=H, R2=OH, was prepared as described in the example 22.
Step 2: A mixture of 10,5-dihydroxycamptothecin of the formula 13 where R2=OH, R1=R3=R4=R5=H (200mg) and trifluoroethanol (1mL) were suspended in 50m1 of dichloroethane and heated to reflex in the presence of sulfuric acid (0.5m1) for 18h. Reaction mixture was concentrated to dryness and the residue was extracted with ethylacetate. Organic layer was washed with water and brine and dried over anh.sodium sulfate.
Evaporation of the solvent furnished an oily residue which was purified over silica gel column using acetonechloroform as an eluent to get 140mg of 10-Hydroxy-5-trifluoroethoxycamptothecin of the formula 1 as a solid; mp: 237°C.; IR: 3420, 1748, 1664, 1605, 1159 cm-1; ZH NMR(DMSO-d6): b 10.48(s, IH, D20 exchangeable), 8.45{s, 1H), 8.04(d, J=9Hz, 1H), 7.47(d, J=9Hz, 1H), 7.40(s, 1H), 7.18(s, 1H), 7.11(s, 0.5H), 7.06(s, 0.5H), 6.58(s, 1H, D20 exchangeable), 5.41(s,2H), 5.05-4.55(m, 2H), 2.05-1.75(m,2H), 1.00-0.8(m,3H); 13C NMR (DMSO-d6) . b 172.4, 161.0, 157.7, 157.1, 151.2, 147.5, 144.3, 143.7, 131.0, 130.8, 129.8, 129.1, 124.0, 121.4, 120.7, 109.6, 96.6, 89.7, 72.3, 65.1, 30.4, 7.8: Mass(m/z): 462(M+1), 418, 364, 320, 291, 263.

Preparation of 9-Nitro-5-trifluoroethoxycamptothecin of the formula 1 where R2=R3=R4-R5=H, Rf=N02 R6=CH2CF3 Step 1: Initially 9-vitro-5-hydroxycamptothecin of the formula 13 where R2=R3=R4=R5=H, RZ=N02 was prepared as described in the example 21.
Step 2: A mixture of 9-vitro-5-dihydroxycamptothecin of the formula 13 where R1=NO2, R2=R3=R4=R5=H (100mg) and trifluoroethanol (0.5mL) were suspended in 25m1 of dichloroethane and heated to reflex in the presence of sulfuric acid (0.3m1) for 18h. Reaction mixture was concentrated to dryness and the residue was extracted with ethylacetate. Organic layer was washed with water and brine and dried over anh.sodium sulfate.
Evaporation of the solvent furnished an oily residue which was purified over silica gel column using WO 97146563 PCTlITS97/06962 acetone-chloroform as an eluent to get 60mg of 9-nitro-5-trifluoroethoxycamptothecin of the formula 1 as a solid.; mp 210°C.; IR: 3457, 1745, 1665, 1623, 1527, 1154, 1000 cm-l; 1H NMR (CDC13) : b 9.30 (s, 1H) , 8.53 (d, J=8.6Hz, 1H) , 8.49 (d, J=8.6Hz, 1H} , 7.94 (t, J=8Hz, 1H) , 7.62(s, 0.5H), 7.60(s, 0.5H), 6.87(s, 0.5H}, 6.81(s, 0.5H), 5.69(d, J=l6Hz, 1H), 5.29(d, J~I6Hz, 1H), 4.97(m, 1H), 4.52(m, 1H), 3.90{br s,lH, D20 exchangeable), 1.90(m,2H), 1.05(t, J=7Hz, 3H); Mass (m/z): 491(M+1), 461, 446, 418, 393, 364, 349, 319, 290, 216 Prevaration of 5-(2'-fluoroethoxy)camptothecin of the formula 1 where RZ=R2=R3=R4=R5=H. Rs=CH2CH2F
Step l: Initially 5-hydroxycamptothecin of the formula 13 where R1=R2=R3=R'~=R5=H, was prepared as described in the example 2.
Step 2: A mixture of 5-hydroxycamptothecin of the formula 13 where R1=R2=R3=R4=R5=H (200mg) and 2-fluoroethanol {2mL) were suspended in 30m1 of dichloroethane and heated to reflex in the presence of sulfuric acid (0.3m1) for 18h. Reaction mixture was concentrated to dryness and the residue was extracted with ethylacetate. Organic layer was washed with water and brine and dried over anh.sodivm sulfate.
Evaporation of the solvent furnished an oily residue which was purified over silica gel column using acetone-chloroform as an eluent to get 130mg of 5-(2'-fluoroethoxy)camptothecin of the formula 1 as a solid;
mp: 174°C.; IR: 1745, 1664, 1615, 1160, 1040, 752 cm-1;
1N NMR {CDC13 + DMSO-d6): b 8.46(s, 1H), 8.20(d, J=8Hz, 1H ), 7.95(d, J=8Hz, 1H), 7.83 (t, 3= 6.8Hz, 1H), 7.65-7.55(m, 2H), 6.86(s, 0.5H), 6.78(s. 0.5H), 5.68(d, ' J=16.5Hz, 1H), 5.26(d, J=16.5Hz, 1H), 4.90-4.20 (m, 4H), 4.44(s, 1H, D20 exchangeable}, 2.05-1.85(m, 2H), 1.12-0.95{m, 3H) ; Mass(m/z): 410(M+1), 365, 348, 319, 304.

Preparation of 10-Hydroxy-5-(2'-fluoroethoxy)-camptothecin of the .formula 1 where R1=R3=R4=R5=H. R2=OH. R6=CH2CH2F
Step 1: Initially 10,5-dihydroxycamptothecin of the formula 13 where R1=R3=R4=R5=H, R2=OH, was prepared as described in the example 22.
Step 2: A mixture of 10,5-dihydroxycamptothecin of the 20 formula 13 where R2=OH, Rl=R3=R'~=R5=H (100mg) and 2-fluoroethanol (2mL) were suspended in 25m1 of dichloroethane and heated to reflex in the presence of sulfuric acid (0.2m1) for 16h. Reaction mixture was concentrated to dryness and the residue was extracted with ethylacetate. Organic layer was washed with water and brine and dried over anh.sodium sulfate.
Evaporation of the solvent furnished an oily residue which was purified over silica gel column using acetone-chloroform as an eluent to get 60mg of 10-Hydroxy-5-fluoroethoxycamptothecin of the formula 1 as a solid; mp: 258- 260°C.; IR: 3225, 1748, 1660, 1593, 1159 cm-1; 1H NMR (CDC13 + DMSO-d6): b 10.0 (br s, 1H, D20 exchangeable), 8.31(s, 1H), 8.00(d, J=6Hz, IH), 7.80(s, 1H), 7.45(d, J=6Hz, 1H), 7.40(s, 1H), 6.85(s, 0.5H), 6.80(s, 0.5H), 6.15(s, 1H, D20 exchangeable), 5.55(d, J=I6Hz, 1H), 5.23(d, J=l6Hz, 1H), 4.85-4.20(m, 4H}, 2.05-1.81(m,2H), 1.0(t, J=7Hz, 3H); Mass(m/z}:
426(Mtl), 382, 364, 320.

Prevaration of 5-(2'-Fluoroethoxy)-7-ethylcamt~tothecin Step 1: Tnitially 5-hydroxy-7-ethylcamptothecin of the ' formula 13 where R~=R2=R3=R4=H, R5=Et, was prepared as described in the example 4.
' Step 2: To a mixture of 80mg of 5-hydroxy-7-ethylcamptothecin and O.lml of p-toluenesulfonic acid suspended a.n l2ml of benzene, 20mg of 2-fluoroethanol was added and heated the mixture to reflex temperature _ 58 _ for 14h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using ethyl acetate-chloroform as eluent to afford 60mg of 5-(2'-fluoroethoxy)-7-ethylcampto-thecin; mp: 112°C.; IR: 3070, 1748, 1665, 1605, 1456, 1155, 1038. 767 cm-1; iH NMR (CDC13): b 8.21 (d, J=9.2Hz, 1H), 8.17(d, J=9.2Hz, 1H), 7.82(t, J=7.4Hz, 1h), 7.67(t, J=7.4Hz, 1H), 7.57(s, 0.5H), 7.54(s, 0.5H), 7.00(s, 0.5H), 6.89(s, 0.5H), 5.69(d, J=l6Hz, 1H), 5.27(d, J=l6Hz, 1H), 4.81-4.12(m, 4H), 3.51-3.15(m, 2H), 1.93(m, 2H), 1.45(t, J=?Hz, 3H), 1.05(m, 3H); Mass(m/z): 438(M+1), 420, 406, 376, 347, 332, 317, 245, 91.

Preparation of 5-(2'-HVdroxyethoxy) 7 ethylcamptothecin Step 1: Initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where R1=R2=R3=R4=H, R5=Et, was prepared as described in the example 4.
Step 2: To a mixture of 250mg of 5-hydroxy-7-ethylcamptothecin and 10 E.cl of-conc.sulfuric acid suspended in 25m1 of dichloroethane, 0.5m1 of ethylene glycol was added and heated the mixture to reflux temperature for 14h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water and brine and concentrated to dryness. The residue was purified by silica gel column chromatography using ethyl acetate-chloroform as eluent to furnish 180mg of 5-(2'-hydroxyethoxy)-7-ethylcamptothecin and 25mg of starting material; 1H NMR
(CDC13, 200MHz): b 8.20(d, J=8Hz, 1H), 8.15(d, J=8Hz, ' 1H), 7.85(t, J=6.8Hz, 1H), 7.69(t, 7.3Hz, 1H), 7.56(s, 0.5H), 7.54(s, 0.5H), 7.11(s, 0.5H), 6.99(s, 0.5H), ' 5.72(d, J=16.5Hz, 1H), 5.28(d, J=16.5Hz, 0.5H), 5.26(d, J=16.5Hz, 0.5H), 3.95-3.65(m, 4H), 3.78 (br s, 2H, D20 exchangeable), 3.5-3.18 (m, 2H), 1.95-1.81(m, 2H), 1.45 (t, J=7 .5Hz, 3H) , 1.06 (m, 3H) .

Preparation of 5-(2'-Hvdroxyethoxy) 10- hydroxycamptothecin Step 1: Initially 10,5-dihydroxycamptothecin of the formula 13 where RZ=R3=R4=R5=H, R2=OH, was prepared as described in the example 22.
Step 2: To a mixture of 60mg of 10,5-dihydroxycamp.to-thecin and 5mg of p-toluenesulfonic acid suspended in lOml of dichloroethane, 25mg of ethylene glycol was added and heated the mixture to reflux temperature for 16h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water and brine and concentrated to dryness. The residue was purified by silica gel column chroma-tography using methanol-chloroform as eluent to furnish 40mg of 5-(2'-hydroxyethoxy)-l0-hydroxycampto-thecin and lOmg of starting material; IR; 3070, 1760, 1660, 1600, 1558, 1509, 1384, 1160, 1047, 832 cm-~; ZH NMR
CDC13 + DMSO): 8 10.05(br, 1H D20 exchangeable), 8.35(s, 1H), 8.05(d, J=9Hz, 1H), 7.75(s, 1H), 7.45(d, J=9Hz, 1H}, 7.28(s, 1H}. 6.95(s, 0.5H), 6.85(s, 0.5H), 5.65(d, J=l6Hz, 1H), 5.25(d, J=l6Hz, 1H), 4.11(m, 2H), 3.78(m, 2H), 4.05(br s, 1H, D20 exchangeable), 1.98(m, 2H}. 1.05(t, J=7Hz, 3H); Mass (m/z): 425, 408, 380, 364, 336, 320, 305, 264, 235, 147, 105.

Preparation of 5,10-Dihydroxy-7-ethylcamntothecin Step 1: Initially 5-ethoxy-10-hydroxy-7-ethylcamptothecin of the formula 1 where R1=R3=R4=H, R5=RS=Et, R2=OH was prepared as described in the Example ~ 18.
Step 2: l2ml of 259sHC1 was added to 200mg of 5-ethoxy-10-hydroxy-7-ethylcamptothecin dissolved in lOml of ethanol and heated to reflux for 24h. Excess acid and ethanol were distilled off and the remaining residue was diluted with ethylacetate. The organic layer was washed with 5o NaHC03 solution and brine.
Concentration of the solvent and purification of the solid material over~silica gel column using acetone-chloroform solvent mixture as eluent afforded 105mg of 5, 10-dihydroxy-7-ethylcamptothecin; mp . 197°C.; IR:
3268, 2975, 1?48, 1656, 1597, 1514, 1230, 1161, 1052, 841 cm-1; 1H NMR (CDC13+DMSO): 8 10.0(br s, 1H, D20 exchangeable), 8.05(d, J=9Hz, 1H), 7.76(s, 2H), 7.42(m, 1H), 7.04(s, 0.5H), 6.98(s, 0.5H), 5.65(d, Jl6Hz, 1H), 5.23(d, J=l6Hz,lH), 3.5I(br s, 1H, D20 exchangeable), 3.45-3.12(m, 2H), 1.94(m, 2H), 1.42(t, J=7Hz, 3H), 1.01(t, J=7Hz, 3H); Mass(m/z): 408{M+1), 379, 364, 347, 335, 285, 169, 119, 101, 83.

Preparation of 5-(2'Hydroxyethoxy) 10 hydroxZr 7-ethYlcamptothecin Step 1: Initially 7-ethyl-5,10-dihydroxy camptothecin of the formula 13 where R1=R3=R4=H, R2=OH, R5=Et, was prepared as described in the example 34.
2Q Step 2: To a mixture of 100mg of 10,5-dihydroxy-7-ethylcamptothecin and 5mg of p-toluenesulfonic acid suspended in lOml of dichloroethane, 50mg of ethylene glycol was added and heated the mixture to reflux temperature for 16h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water and brine and concentrated to dryness. The residue was purified by silica gel column chromatography using methanolchloroform as elueat to furnish 60mg of 5-{2'-hydroxyethoxy)-10-hydroxy-7-ethyleamptothecin and l2mg of starting material.; mp: 124°C.; 1H NMR (CDC13+DMS~:~: 8 10.0 (br s, 1H, D20 exchangeable), 8.02(d, J=9Hz, 1H), 7.55- ' 7.39(m, 3H), 7.02(s, 0.5H), 6.93(s, 0.5H), 6.05 (br s,lH, D20 exchangeable), 5.63(d, J=l6Hz, 1H}, 5.23(d, J=l6Hz, IH), 3.94-3.54(m,2H), 3.41-3.05(m 2H), 1. 93 (m, 2H) , 1.40 (t, J=7Hz, 3H) , 1. 02 (m, 3H) ; Mass (m/z) 408(M+1), 379, 364, 347, 335, 285, 169, 119, 101, 83.

_ 61 _ Preparation of 5-(2'-aminoethoxy) camptothecin Step 1: Initially 5-hydroxy camptothecin of the formula 3.3 where R1=R2=R3=R4=R5=H, was prepared as described in the example 2.
Step 2: To a mixture of 60mg of 5-hydroxycamptothecin and 5mg of p-toluenesulfonic acid suspended in l0ml of benzene, l5mg of 2-aminoethanol was added and heated the cure to reflux temperature for 14h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using methanolchloroform as eluent to furnish 36mg of 5-(2'-aminoethoxy)camptothecin and l0mg of starting material;
mp: 170°C.; IR: 3451, 1740, 1664, 1604, 1383, 1189, 1042 cm-1; Partial 1H NMR data in (CDC13 + DMSO-d6): b 7.5(d, D20 exchangeable, 1H), 7.15(d, D20 exchangeable, 1H), 7.02 (s, 0.5H), 6.92(s, 0.5H), 5.65(d, J=l6Hz, 1H), 5.28(d, J=l6Hz, 1H), 4.24-3.85(m, 2H), 2.35(s, D20 exchangeable, 1H), 2.34(m, 1H), 2.15-1.85(m, 3H). 1.12-0.95(m, 3H); Mass (m/z): 408(M+1), 364, 347, 319, 305, 291, 249, 103, 62.

Prer~aration of 5- (2' -aminoethoxy) -7-ethylcamQtothecin Step 1: Initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where R1=R2=R3=R4=H, R5=Et, was prepared as described in the example 4.
Step 2: To a mixture of 85mg of 7-ethyl-5-hydroxycamptothecin and 5mg of p-toluenesulfonic acid suspended a.n 20m1 of benzene, 11 mg of 2-aminoethanol was added and heated the mixture to reflux temperature for lOh. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using methanol-chloroform as eluent to WO 97/46563 PC~'/I1S97/06962 afford 65mg of 5-(2'-aminoethoxy)-7-ethylcamptothecin.
mp: 230°C.; Partial 1H NMR in (CDC13+ DMSO-d6): 8 7.5(d, D20 exchangeable, 1H), 7.15(d, D20 exchangeable, 1H) , 7 .02 (s, 0.5H) , 6.92 (s, 0.5H) , 5.65 (d, J=l6Hz, 1H) , 5.28(d, J=l6Hz, 1H), 4.24-3.85(m, 2H), 2.35(s, D20 exchangeable, 1H), 2.34(m, 1H), 2.15-1.85(m, 3H), 1.12-0.95(m, 2H); Mass (m/z): 408(M+1), 364, 347, 319, 305, 103, 74, 62.

Preparation of 5-(3'-dimethylaminopropoxy)-7-ethylcamt~tothecin Step l: Initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where RZ=R2=R3=R~=H, R5=Et, was prepared as described in the example 4.
Step 2: To a mixture of 50mg of 7-ethyl-5-hydroxycamptothecin and 0.05m1 of sulfuric acid suspended in 20m1 of benzene. 30mg of 3-dimethylamino-1-propanol was added and heated the mixture to reflux temperature for 12h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using methanol-chloroform as eluent to obtain 42mg of 5-(3'-dimethylaminapropoxy)-7-ethylcamptothecin; mp: 113°C.;
Partial 1H NMR data in (CDC13+ DMSO-d6): b 6.95 (s, 0.5H), 6.85(s, 0.5H), 5.65(d, J=l6Hz, 1H), 5.35(d, J=l6Hz, 0.5H), 5.25(d, J=l6Hz. 0.5H), 3.95-3.57(m, 2H), 3.30-3.05(m. 2H), 2.85(s, 3H), 2.83(s, 3H), 2.15-1.72(m, 6H), 1.45(t, J=7.5Hz, 3H), 1.12-0.95(m, 3H);
Mass (m/z): 478(M+1), 434, 375, 347, 331, 169, 102, 84;
Mass(m/z) . 478(M+1), 434, 375, 347, 331, 169, 102, 84.

Preparation of 5-(2'-N-rwrrolidinoethoxy)-7-ethvlcamptothecin Step 1: Initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where R1=R2=R3=R4=H, R5=Et, was prepared as described in the example 4.
Step 2: To a mixture of 100mg of 7-ethyl-5-hydroxycamptothecin and lOmg of camphorsulfonic acid suspended in 25m1 of benzene, 30mg of 1-(2-hydroxyethyl) pyrrolidine was added and heated the mixture to reflex temperature for 16h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using methanol-chloroform as eluent to acetate 85mg of 5-(2'-N-pyrrolidinoethoxy)-7-ethylcamptothecin; mp: 225°C.;
IR: 3424, 1749, 1666, 1616, 1384, 1156, 1078, 1049 em-1; Partial 1H NMR data in CDC13: d 7.02 (s, 0.5H), 6.95(s, 0.5H), 5.70(d, J=l6Hz, 1H), 5.33(d, J=l6Hz, 0.5H), 5.26(d, J=l6Hz, 0.5H), 4.18-3.88(m, 2H), 3.45-3.15(m,2H), 3.06-2.58(m, 6H), 2.05-1.72(m, 6H), 1.43(t, J=8Hz, 3H), 1.15-0.95(m,3H); Mass (m/z): 446 (M+1);
375, 347, 331, 245, 169, 116, 97, 84.

Preparation of 5-(2'-chloroethoxy)-7-ethylcamptothecin Step 1: Initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where R1=R2=R3=R4=H, R5=Et, was prepared as described in the example 4.
Step 2: To a mixture of 500mg of 7-ethyl-5-hydroxycamptothecin and O.lml of conc.sulfuric acid suspended in 30m1 of benzene, 700mg of 2-chloroethanol was added and heated the mixture to reflex temperature using Dean-Stork apparatus for 8h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using ethyl acetate-chloroform as eluent to provide 400mg of 5-(2'-chloroethoxy)-7-ethylcamptothecin; mp: 168°C.; 1H
NMR (CDC13, 200MHz) : S 8.20 (d, J=9.5Hz, 1H) , 8.15 (d, J=9.5Hz, 1H), 7.82(t, J=8Hz, 1H}, 7.67(t, J=8Hz, 1H), 7.54(d, 6.2Hz, 1H), 7.02(s, 0.5H), 6.90(s, 0. 5H), 5.70(d, J=l6Hz, 0.5H), 5.69(d, J=l6Hz, 0.5H), 5.26{d, J=l6Hz, 0.5H), 5.25(d, J=l6Hz, 0.5H), 4.61-3.95(m, 2H), 3.78-3.58{m, 2H), 3.50-3.15(m, 2H), 1.98-1.78(m, 2H), 1.45-(t, 3=7. SHz, 3H), 1.12-0.95(m, 3H}; Mass (m/z):
455(Mtl), 437, 409, 392, 376, 347, 331, 245, 115, 81.

Pret~aration of 5-(2'-dimethylaminoethoxy)-7-ethylcamptothecin Step 1: initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where Rl=R2=R3=R4=H, R5=Et, was prepared as described in the example 4.
Step 2: To a mixture of 100mg of 7-ethyl-5-hydroxycamptothecin and 0.lml of conc.sulfuric acid suspended in 10m1 of benzene, 50mg of 2-N,N-dimethylaminoethanol was added and heated the mixture to reflux temperature using Dean-Stark apparatus for 10h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness.
The residue was purified by silica gel column chromatography using methanol-chloroform as eluent to get 65mg of 5-(2'-dimethylaminoethoxy)-7-ethylcamptothecin; Partial 1H NMR data in CDC13: S 7.05 (s, 0 .5H) , 6.93 (s, 0 .5H) , 5.74 (d, J=I6Hz, 0 .5H) , 5.73(d, J=l6Hz, 0.5H), 5.29(d, J=l6Hz, 1H), 4.41-3.75(m, 2H), 3.53-3.18(m, 2H), 2.57(q, J=6Hz, 2H), 2.26(s, 3H), 2.23{s, 3H), 2.05-1.86(m, 2H), 1.47(t, J=SHz, 3H), 1.18-1.02(m, 3H).

Preparation of 5-(4'-aminobutoxy) camptothecin Step 1: Initially 5-hydroxy camptothecin of the formula 13 where R1=RZ=R3=R4=R5=H, was prepared as ' described in the example 2.
Step 2: To a mixture of 53mg of 5-hydroxycamptothecin and 8mg of p-toluenesulfonic acid suspended in 16m1 of WO 97!46563 PCT/US97/06962 benzene, l4mg of 4-aminobutanol was added and heated the mixture to reflux temperature for 10h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using ethyl acetatechloroform as eluent to furnish 45mg of 5-(4-aminobutoxy) camptothecin; mp: 150°C.: IR: 3397, 1745, 1664, 1617, 1384, 1224, 1162, 1038, 684, 570 cm-~; Partial 1H NMR data in (CDC33+ DMSO-d6): b 7.50(d, D20 exchangeable, 1H), 6.95(s, 0.5H), 6.85(s, 0.5H), 6.25(d, Da0 exchangeable, 1H), 5.65(d, J=l6Hz, 1H), 5.35(d, J=l6Hz, 0.5H), 5.25(d, J=l6Hz, 0.5H), 4.15-3.80(m, 2H), 2.15-1.65(m, 8H), 1.15-0.98(m, 3H); Mass (m/z): 436(M+1). 392, 347, 333, 305, 153, 123, 105, 90, 62;

Preparation of 5-(2'-metho~ethoxy)camptothecin Step 1: Initially 5-hydroxy camptothecin of the formula 13 where R1=R2=R3=R4=R5=H, was prepared as described in the example 2.
Step 2: To a mixture of 120mg of 5-hydroxycamptothecin and 0.13m1 of sulfuric acid suspended in 18m1 of chloroform, 20mg of ethyleneglycol monomethylether was added and heated the mixture to reflux temperature for lOh. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate. Organic layer was washed with water, NaHC03, brine and concentrated to dryness.
The residue was purified by silica gel column chromatography using ethyl acetate-chloroform as eluent to furnish 80mg of 5-(2'-methoxyethoxy)camptothecin;
mp: 123°C.; IR, 3294, 2933, 1748, 1665, 1617, 1384, 1155, 1077, 1045, 761 cm-1; 1H NMR (CDC13): b 8.51(s, 1H) , 8.24 (d, J=8Hz, 1H) , 7. 93 (d, J=8Hz, 1H) , 7.79 (t, J=6.8Hz, 1H), 7.65(t, J=6.8Hz, 1H), 7.58(s, 0.5H), 7.56(s, 0.5H), 6.91(x, 0.5H), 6.82(s, 0.5H), 5.71(d, J=l6Hz, 1H), 5.28(d, J=l6Hz, 1H), 4.55-4.05(m,2H), 3.95(br s, 1H, D20 exchangeable), 3.81-3.56(m,2H), 3.48(s, 1.5H), 3.44(s, 1.5H), 1.94 (m,2H), 1.05(t, J=7Hz, 3H); Mass(m/z):~423(M+1), 364, 347, 319, 304, 275, 218, 128, 101, 82.

Preparation of 5-(2'N-Methyltwrrolidinoethoxy)-7-ethylcamptothecin Step 1: Initially 5-hydroxy-7-ethylcamptothecin of the formula 13 where Rz=R2=R3=R4=H, R5=Et, was prepared as described a.n the example 4.
Step 2: To a mixture of 50mg of 5-hydroxy-7-ethylcamptothecin and l0mg of p-toluenesulfonic acid suspended a.n 15m1 of benzene, l8mg of 1-methyl-2-pyrrolidinoethanol was added and heated the mixture to reflex temperature for 8h. Reaction was quenched with a drop of pyridine and extracted with ethyl acetate.
Organic layer was washed with water, NaHC03, brine and concentrated to dryness. The residue was purified by silica gel column chromatography using methanol-chloroform as eluent to furnish 35mg of 5-(2'-N-Methylpyrrolidinoethoxy)-7-ethylcamptothecin; mp:
102°C.: Mass (m/z): 504(M+1), 460, 375, 347, 331, 275, 245, 128, 110, 84.

Claims (13)

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A compound of formula 1, or its pharmaceutically acceptable salts, wherein R1, R2, R3, and R4 independently represent hydrogen or represent a group selected from hydroxy, (C1-C8)alkoxy, nitro, substituted amino where the amino group is mono or disubstituted wherein the substituent is selected from (C1-C8)alkyl, halo(C1-C 8)alkyl, or substituted (C1-C8)alkyl, wherein the substituents are selected from hydroxy, (C1-C8)alkoxy, amino, or (C1-C8) alkylamino;
R5 represents hydrogen, or (C1-C8)alkyl ; and R6 represents hydrogen, (C1-C8)alkyl, benzyl where the phenyl group may be unsubstituted or substituted with mono, di or trisubstituents selected from halogen, (C1-C8)alkoxy, (C1-C8)alkyl, amino, or substituted amino where the amino group is mono or disubstituted with (C1-C8)alkyl group; (C1-C8)alkyl groups substituted with a heterocyclic ring having 3 to 7 ring atoms said ring atoms being carbon atoms and one or two heteroatoms said heteroatoms selected from oxygen, nitrogen or sulfur;
substituted (C1-C8)alkyl, where the substituents are selected from halogen, hydroxy, (C1-C8)alkoxy, aryloxy, pyridyl, cyano, nitro, or amino in which the amino group can be unsubstituted or mono, or disubstituted wherein the substituent is selected from hydroxy, or (C1-C8) alkyl, when the amino group is disubstituted the substituents are independent or combined together to form a cyclic ring having 5 or 6 ring atoms the ring atoms are carbon atoms, nitrogen atom and optionally one or two heteroatoms selected from nitrogen or oxygen;
with the provisos that (i) when R1 is methoxy, R6 is not hydrogen or unsubstituted (C1-C8)alkyl group; (ii) when R2 is hydroxy, (C1-C8)alkoxy, nitro, amino, alkylamino or halogen, R6 is not hydrogen or unsubstituted alkyl group; (iii) when R5 is (C1-C8)alkyl, R6 is not hydrogen or (C1-C8)alkyl group; (iv) when R1 is methoxy group, R2 is hydroxy, (C1-C8)alkoxy, nitro, amino; alkylamino or halogen, R5 is (C1-C8)alkyl, R6 is not hydrogen or unsubstituted (C1-C8) alkyl group and (v) when R1-R5 represent hydrogen, R6 is not hydrogen or unsubstituted (C1-C8)alkyl group.

2. A compound according to claim 1, selected from:
5-trifluoroethoxy CPT;
9-nitro-5-(2'-methoxyethoxy) CPT;
9-hydroxy-5-ethoxy CPT;
9-nitro-5-ethoxy CPT;
7-Ethyl-5-(2-chloroethoxy) CPT;
5-(2'-hydroxyethoxy) CPT;
10-hydroxy-5-(2'-hydroxyethoxy) CPT;
7-ethyl-10-hydroxy-5-(2'-hydroxyethoxy) CPT;
9-nitro-5-fluoroethoxy CPT;
10-hydroxy-5-trifluoroethoxy CPT;
7-ethyl-10-hydroxy-5-trifluoroethoxy CPT;
7-ethyl-5-pyrrolidinoethoxy CPT;
7-ethyl-5-dimethylaminopropoxy CPT;
7-ethyl-10-hydroxy 5-fluoroethoxy CPT;
5-(2'-hydroxyethoxy)-7-ethyl CPT and 5-(2'-methoxyethoxy) CPT
where CPT represents 20(S)-camptothecin.

3. Compounds of formula 1 where R1 through R6 have the meaning defined in any one of claims 1 to 2 as a mixture of two diastereomers, said diastereomers having 20(S), 5(R) and 20(S), 5(S), configurations.

4. Compounds of formula 1 in which each of the diastereomers having 20(S),

5(R) and 20(S), 5(S) configurations as individual isomer, substantially free from the other isomer, where R1 through R6 have the meaning defined in any one of claims 1 to 2.
6. A pharmaceutical composition comprising a compound of formula I as defined in any one of the claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable non-toxic excipient, diluent or solvent.

6. A compound of formula 1 as defined in any one of claims 1 to 4 for the treatment of cancers, leukemia or HIV related conditions.

7. Use of a compound according to any one of claims 1 to 4 for treatment of cancer, leukemia or HIV related conditions.

8. A process for the preparation of a compound of formula 1, which comprises the steps of:
(i) reacting a compound of formula 2, wherein R1, R2, R3, and R4 independently represent hydrogen or represent a group selected from hydroxy, (C1-C8)alkoxy, nitro, substituted amino where the amino group is mono or disubstituted wherein the substituent is selected from (C1-C8)alkyl, halo(C1-C8)alkyl, or substituted (C1-C8)alkyl, wherein the substituents are selected from hydroxy, (C1-C8)alkoxy, amino, or (C1-C8) alkylamino; and R5 represents hydrogen, or (C1-C8)alkyl, in the presence of acid said acid selected from inorganic acid or Lewis acids, and a ferric salt, with a compound having the formula R6 -OH where R6 represents (C1-C8)alkyl, (C1-C8)alkenyl, (C3-C7)cycloalkyl, haloalkyl or hydroxyalkyl to obtain a compound of the formula 12 and a compound of the formula 13, wherein R1, R2, R3, R4 and R5 have the meaning given above, optionally;
(ii) separating the compounds of the formula 12 and 13 prepared in step (i), optionally;
(iii) hydrolyzing the compound of the formula 12 to obtain additional amounts of the compound of the formula 13, and optionally, (iv) reacting the compound of the formula 13, in the presence of an acid selected from inorganic acids, Lewis acids or organic acids with a compound having the formula R6-OH to obtain a compound of the formula 1, wherein R1, R2, R3, and R4 independently represent hydrogen or represent a group selected from hydroxy, C1-C8)alkoxy, nitro, substituted amino where the amino group is mono or disubstituted wherein the substituent is selected from (C1-C8 )alkyl, halo (C1-C8)alkyl, wherein the substituents are selected from hydroxy, (C1-C8)alkoxy, amino, or (C1-C8)alkylamino;
R5 represents hydrogen or (C1-C8)alkyl, and R6 is (C1-C8)alkyl, benzyl where the phenyl group may be unsubstituted or substituted with mono, di or trisubstituents selected from halogen,(C1-C8)alkoxy, cyano, nitro, (C1-C8)alkyl, amino, or substituted amino where the amino group is mono or disubstituted with (C1-C8)alkyl group; (C1-C8) alkyl groups substituted with a heterocyclic ring having 3 to 7 ring atoms said ring atoms being carbon atoms and one or two heteroatoms said heteroatoms selected from oxygen, nitrogen or sulfur; substituted (C1-C8)alkyl where the substituents are selected from halogen, hydroxy, (C,-C8)alkoxy, aryloxy, pyridyl, cyano, nitro, or amino in which the amino group can be unsubstituted or mono, or disubstituted wherein the substituent is selected from hydroxy, or (C1-C8)alkyl, when the amino group is disubstituted the substituents are independent or combined together to form a cyclic ring having 5 or 6 ring atoms the ring atoms are carbon atoms, nitrogen atom and optionally one or two heteroatoms selected from nitrogen or oxygen;
with the provisos that (i) when R1 is methoxy, R6 is not hydrogen or unsubstituted (C1-C8)alkyl group; (ii) when R2 is hydroxy, (C1-C8)alkoxy, nitro, amino, alkylamino or halogen, R6 is not hydrogen or unsubstituted alkyl group; (iii) when R5 is (C1-C8)alkyl, R6 is not hydrogen or (C1-C8)alkyl group; (iv) when R1 is methoxy group, R2 is hydroxy, (C1-C8)alkoxy, nitro, amino, alkylamino or halogen, R5 is (C1-C8)alkyl, R6 is not hydrogen or unsubstituted (C1-C8) alkyl group and (v) when R1-R5 represent hydrogen, R6 is not hydrogen or unsubstituted (C1-C8)alkyl group.

9. A process according to claim 8, for the preparation of a compound of formula 1, where R1, R3, R4 and R5 are hydrogen, R2 represents hydroxyl group and R6 represents trifluoroethyl group, which comprises the steps of:
(i) reacting a compound of the formula 2, where R1, R3, R4 and R5 are hydrogen, and R2 represents hydroxyl group, in the presence of concentrated sulfuric acid and ferric chloride trihydrate with ethanol and heating the mixture to reflux conditions to obtain a compound of the formula 12 and compound of the formula 13, wherein R1, R3, R4 and R5 represent hydrogen, and R2 is hydroxyl group and R6 represents ethyl group, (ii) separating the compounds of formula 12 and 13 prepared in step (i), (iii) hydrolyzing the compound of the formula 12, by dissolving in aqueous ethanol and refluxing with hydrochloric acid, to obtain additional amounts of the compound of formula 13, (iv) reacting the compound of the formula 13, in the presence of conc.sulfuric acid, with trifluoroethanol dissolved in dichloroethane solvent, to obtain the compound of the formula I, where R1, R3, R4 and R5 are hydrogen, R2 represents hydroxyl group and R6 represents trifluoroethyl group.

10. A process for the preparation of compounds of formula 1, wherein R6 represents hydrogen or (C1-C8)alkyl, R1 represents hydrogen or methoxy, R2 represents hydrogen, hydroxy, (C1-C8)alkoxy, acyloxy, SH, thioalkyl, thioacyl, nitro, amino, alkylamino, acylamino or halogen; R3 and R4 are hydrogen and R5 represents hydrogen, (C1-C8)alkyl, (C1-C8)aralkyl, CH2OH, COOH, COOMe or CH2OR' where R' represents (C1-C8)alkyl or acyl group which comprises, the steps of:
(i) reacting a compound of the formula 2, where R1 to R5 have the meaning described above, in the presence of an acid selected from inorganic acids, or Lewis acids and a ferric salt, with a compound of the formula R6-OH
where R6 represents a (C1-C8)alkyl group, to obtain a compound of the formula and a compound of the formula 13, wherein R1, R2, R3, R4 and R5 have the meaning given above, optionally (ii) separating the compounds of formula 12 and 13 prepared in step (i), (iii) hydrolyzing the compounds of the formula 12 to obtain additional amounts of the compound of the formula 13, and optionally, (iv) reacting the compound of the formula 13, in the presence of an acid, with a compound having the formula R6-OH to obtain compounds of the formula 1, where R1, R2, R3, R4 and R5 have the meaning described above, and R6 represents a (C1-C8) alkyl group.

11. The process according to claim 8 or 10, wherein the ferric salt is ferric chloride.

12. A pharmaceutical composition according to claim 5, that is formulated for the treatment of cancer, leukemia, or an HIV-related condition.

13. Use of a compound according to any one of claims 1 to 4, for the manufacture of a medicament for the treatment of cancer, leukemia or an HIV-related condition.