J. Comb. Chem. 2003, 5, 653-659
653
Solution-Phase Synthesis of a Library of 3,5,7-Trisubstituted 3H-[1,2,3]triazolo[4,5-d]pyrimidines Nand Baindur, Naresh Chadha, and Mark R. Player* 3-Dimensional Pharmaceuticals Inc., 8 Clarke DriVe, Cranbury, New Jersey 08512 ReceiVed December 2, 2002 An efficient solution-phase parallel synthesis of a library of 3,5,7-trisubstituted [1,2,3]triazolo[4,5-d]pyrimidines is described. Monosubstituted amidines may be converted to 2-substituted 5-amino-4,6dihydroxypyrimidines in four steps. Treatment with a primary amine followed by cyclization yields the 7-chloro-3,5-disubstituted [1,2,3]triazolo[4,5-d]pyrimidines as penultimate intermediates. Final nucleophilic substitution of the 7-chloro group with an excess of a primary or secondary amine, a hydrazine or a O-alkylhydroxylamine proceeds efficiently. Scavenging of the excess amine with a resin-bound isocyanate in the presence of resin-bound piperidine as a base affords the desired 3,5,7-trisubstituted [1,2,3]triazolo[4,5-d]pyrimidines in good yields and purities. Introduction In recent years, combinatorial chemistry has emerged as a major tool for accelerating drug discovery through its impact on both lead generation and lead optimization strategies,1 and both solid- and solution-phase parallel synthetic approaches have been adopted for this purpose.1,2 In particular, solution-phase strategies have become increasingly popular, since traditional analytical techniques such as TLC, HPLC (ELSD), and NMR can be readily utilized to monitor reactions.2 Moreover, reactions can sometimes be carried out in a wider variety of solvents and under more stringent conditions.2 Owing to the important role played by naturally occurring purines in several biochemical and physiological pathways, purine derivatives and their various isosteric analogues have been of interest over the years for a variety of biological targets and therapeutic indications. More recently, purine derivatives, such as olomoucine, purvalanol, roscovitine, and related analogues, have been identified as potent inhibitors of kinases, while other purine derivatives, such as myoseverin, have been identified as potent inhibitors of microtubule assembly processes.3 In view of these findings, purines and related structural classes may be privileged scaffolds for drug discovery and high throughput screening. 3,5,7-Trisubstituted [1,2,3]triazolo[4,5-d]pyrimidines, which can be considered as 8-aza analogues of similarly substituted purines (Scheme 1), are one such example.4 We have developed a facile and rapid solution-phase parallel approach for a diverse library of these compounds. [1,2,3]Triazolo[4,5-d]pyrimidines can in principle be synthesized (Scheme 2) by either a triazole-first approach (assembly of a suitably substituted triazole followed by cyclization) or the pyrimidine-first approach (assembly of a suitable substituted pyrimidine followed by cyclization). Both * To whom correspondence should be addressed. Phone: 609-655-6950. Email:
[email protected].
Scheme 1
approaches have been described in the literature, with the triazole-first approach particularly described in the published literature5 and the pyrimidine-first approach described in the patent literature.6 However, there has not been any published or patented approach to readily generate libraries of compounds in this series using either of the two approaches described above. We therefore set out to develop a general method to prepare a library of compounds using the pyrimidine-first approach. Results and Discussion The synthetic scheme for the pyrimidine-first approach is depicted in Scheme 3. The first point of diversity (R1) is introduced using an amidine. A N-unsubstituted amidine (1) is condensed with diethylmalonate (2) in the presence of sodium ethoxide to obtain the 2-substituted 4,6-dihydroxypyrimidine (3). Controlled nitration of (3) with fuming nitric acid gives the corresponding 5-nitro analogue (4). Chlorination with phosphorus oxychloride results in complete conversion of the dihydroxypyrimidines to the corresponding dichloropyrimidines (5) in good yields. The dichloronitropyrimidines are then reduced to the corresponding dichloroaminopyrimidines (6) with iron and acetic acid. The mild conditions for this reduction are necessary to prevent reductive dechlorination of one or both of the chlorines on the pyrimidine ring. At this stage, the second point of diversity (R2) is introduced into the molecule. Thus, 2-substituted 5-amino-4,6-dichloro-pyrimidines are alkylated by
10.1021/cc020110x CCC: $25.00 © 2003 American Chemical Society Published on Web 07/18/2003
654 Journal of Combinatorial Chemistry, 2003, Vol. 5, No. 5
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Scheme 2
Scheme 3a
a
(a) EtONa/EtOH; (b) HNO3; (c) POCl3, PhN(Et)2; (d) Fe/AcOH; (e) R2-NH2; (f) NaNO2/AcOH/DCM; (g) R3-NH2(excess); PS-piperidine/PS-isocyanate.
heating with substituted primary amines, including aromatic (substituted anilines) and aliphatic amines (benzylamine) for 24-48 h at high temperature. This predominantly results in monoalkylation of the symmetric molecules (7). Some dialkylated product formation occurs as well, but chromatographic purification at this stage is rarely required. The monoalkylated product is then readily cyclized with sodium nitrite using mild acidic conditions to generate the triazolopyrimidines (8) in good yields. Purification by flash chromatography is necessary at this stage because it is the penultimate stage in the synthesis. The cyclized products are relatively nonpolar and are readily separated from the more polar impurities using parallel silica gel flash chromatography. Once all the intermediates (8) are obtained, the final point of diversity (R3) is introduced in the molecules via the nucleophilic displacement of the last remaining chlorine with a variety of amines (primary and secondary amines, aromatic amines, hydrazines, and hydroxylamines) to yield the desired
3,5,7-trisubstituted triazolopyrimidines (9). This step can also be considered as a “diversity explosion” step, because each intermediate (8) can be treated with a diverse set of amines, hydrazines, or O-alkylhydroxylamines at this stage to generate a fairly large library of diverse compounds. Purification of the final compounds is unnecessary, because the reactions are driven to completion by use of an excess of the amines followed by scavenging of the excess amine with a suitable solid-phase scavenger (PS-isocyanate) in the presence of a polymer-bound base (PS-piperidine). Parallel filtration and washing of the resins followed by parallel concentration of the filtrates gives the desired 3,5,7-trisubstituted 3H-[1,2,3]triazolo[4,5-d]pyrimidines in good yields and purities (Table 1). In general, both primary amines and secondary aliphatic amines appear to give good yields (57-100%) and purities (63-100%). However, a less nucleophilic aromatic amine, such as aniline, under the same conditions does not always
Synthesis of Pyrimidines
Journal of Combinatorial Chemistry, 2003, Vol. 5, No. 5 655
Table 1. MS and HPLC (ELSD) Purities of Library Compounds Synthesized
entry no. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64
cmpd no.
9i 9j 9k 9l 9m 9n 9o 9p 9q
9t
R1
R2
R3 amine used
MW
mass found
purity (%)
H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H H methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl
benzyl benzyl benzyl benzyl benzyl benzyl benzyl benzyl phenyl phenyl phenyl phenyl phenyl phenyl phenyl phenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl benzyl benzyl benzyl benzyl benzyl benzyl benzyl benzyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-chlorophenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl p-methoxyphenyl
benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine
316 270 302 308 317 332 395 390 302 256 288 294 303 318 381 376 332 286 318 324 333 348 411 406 337 291 323 329 338 353 416 411 316 270 302 308 317 332 395 390 330 284 316 322 331 346 409 404 351 305 336 343 352 367 430 425 346 300 332 338 347 362 425 420
317 271 303 309 318 333 396 391 303 257 289 295 304 319 382 377 333 287 319 325 334 349 412 407 338 292 324 330 339 354 417 412 317 271 303 309 318 333 396 391 331 285 317 323 332 347 410 405 352 306 337 344 353 368 431 426 347 301 333 339 348 363 426 421
94.6 98.5 93.2 99.6 95 99.4 98.9 87.1 98.7 85.9 94.5 99.2 64 99.6 98.6 98.8 96 97 85.8 97.3 68 93.1 96.1 95 95 96.1 87 98.8 90.9 95.7 95.5 98.5 100 99.1 99.2 100 92.8 100 100 99.8 94 96.8 89.9 99.5 69 99 98.4 90 100 99 99.6 100 93 100 100 100 96.1 76 86 99.5 78 98.4 97.4 93.7
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Table 1. (Continued) entry no.
cmpd no.
R1
R2
R3 amine used
MW
mass found
purity (%)
65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80
9a 9b 9c 9d 9e 9f 9g 9h 9r
methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl methyl
phenyl phenyl phenyl phenyl phenyl phenyl phenyl phenyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl p-tolyl
benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine benzyl ethanolamine aniline cyclohexylamine phenylhydrazine benzyloxyamine Boc-piperazine homoveratrylamine
316 270 302 308 317 332 395 390 330 284 316 322 331 346 409 404
317 271 303 309 318 333 396 391 331 285 317 323 332 347 410 405
100 87.6 98.9 100 89.9 100 100 96.4 100 66 94.6 96.6 87.2 94.5 100 95.3
9s
yield as high a purity (see, for example, entries 19, 27, 43, and 59 in Table 1, which show 90% purities). The same is true of a hydrazine, such as phenylhydrazine which tends to give less than desirable purities in several cases (see, for example, entries 13, 21, 45, 61, 69, and 77 which show 90% purities). Finally, the scope of this methodology extends to amines containing an unprotected hydroxyl group, as well, although the purities may be somewhat less than desired. Thus, ethanolamine with an unprotected hydroxyl group gives only moderately good purities in several instances (for example, entries 10, 58, 66, and 74 show
