CA1289138C - Intermediates for purine derivatives - Google Patents

Abstract

ABSTRACT
Intermediates for novel purine derivatives, particularly novel guanines and hypoxanthines, are described as agents for treating autoimmune diseases. Also novel methods of manufacture for the intermediates.

Description

This is a divisional application of copending application, ; , Serial No. 492,727,, filed Octob~r 10, 1985.

BACKGROUND OF THE INrVE~TION

8-Aminoguanine, a compound known since the turn of the century, has been reported to have PNP-activity by R. ParXs, et. al., in Biochem. Pharm., 31 (2), 163 (1982).
~-(2-Furfuryl)guanine is a known compound described in ~. Am. C~.e~. Soc., 81, 3046 (1959) having no disclosed utility. The present invention is related to novel purine derivatives not obvious to an ordinarily skilled artisan, particularly, 9-heteroaryl guanines as having PNP-inhibiting activity.
8-Amino-9-benzylguanine was discussed at the 16th Annual Graduate Student Meetiny in Medicinal Chemistry, University of Michigan, Ann Arbor, Michigan.
However, the present compounds are not obvious from either the synthesis or biological activity of 8-amino-9-~enzylquanine discussed.
With regard to various novel processes of the present invention Ji-Wang Chern, et al, describe "A
Convenient Synthesis of 2-N-methoxycarbonyl-aminooxazolo[5,4-d]pyrimidines" in_J. Het. Chem. 21, 1245~6 (1984). A similar synthesis is described by S. Ram, et al, in "A Synt~esis of Carbamic AcidCImidazo-Heteroaromatic]Methyl Ester Derivatives Using Methoxycar~onyl Isothiocynate," eterocycles, Vol. 22, ~o. 8, 1984, pp 1789-90, in which methoxy-carbonyl isothiocyanate is ~sed in a one pot reagent for the ring closure of an o-diaminopyrimidine derivativs to afford a purine derivative possessins the metho~ycarbonylamino unctionali~y at position eight. -Fl~rther, the mechanism of these two syn-t-nesis is disc~ssed ~y Ji-Wang C~e-n, et al, in "The ~ovel Rins Opening of an Oxa~olo~5,4-d]Pyrimidine and Subsequent Rearr~ngemen~ to ~orm an Imidazo~4,5-d]
35 Pyrimidine," ~eter?cycles, Vol. 22, ~o. 11, 1984, pp 2439-2441.

2~ B
~DZ-l -3~
None of the disclosures include a disclosure of reaction conditions, or an Ar as a heteroaryl or substituted heteroaryl, substituent defined hereinafter for the compound of Formula I prepared by the novel processes of the present invention. That i5 t corresponding Ar groups as defined hereinafter for each of the novel intermediates III, II, and I to be heteroaryl or substituted heteroaryl are not included in the ab~V~ references and furthermore are not obvious variants thereof.

SU~MARY OF THr Ii~VEI~TIOL~

The present invention relates to a compound of the formula Rl ¦ I
J~

wherein Rl is ~H or sa; R2 is hydrogen, ~HR in whicn ~ is hydrogen or ~OR6-where R6 is al~yl of one to four carbon atoms~ aryl or-arylalKyli ~3 is hydrogen, hydroxyl, mercaoto, bromine or ~H~ whe-e K
is hydrogen or COR6 wherein ~6 is as defined above;
n is zero or one; m is zero, one, two, or three, with tne proviso that m or n is at least one; ~ and Rs are each independently hydrogen, alkyl of one to four carbon atoms, hydroxyal~yl of one to four carbo~.
atoms, aryl, arylal~yl or cycloal~yl of three to six carbon atoms, and Ar is heteroaryl or hetero~ryl , ~2~
~Dz-~l substituted by alkyl, alkoxy of one to four carbon atoms, -C=C-C=C- attached to adjacent carbons so as to form a benzo radical, or halogen; or a pharmaceutically acceptable acid or base addition salt thereof, excluding the compound wherein ~1 is OH, R2 is amino, ~3 is hydrogenf n is zero, m is one, and Ar is 2-furanyl, i.e., 9-(2-furanylmethyl)guanine.
The present invention inclucles a method of manufacture and a pharmaceutical composition comprising an effective amount of a compound of the For~ula I with a pharmaceutically acceptable carrier, as well as a method of treatment of autoimmune diseases such as arthritis, systemic lupus erythematosus, inflammatory bo~el diseases, multi~le lS sclerosis, juvenile diabetes, as well as transplantation, viral infections and cancer by administering an effective amount of a compound of the Formula I in unit dosage form to a host of the disease.
That is, the amount is the amount effective for 2~ treating each of the autoimmune diseases. It is understood, an ordinarily skilled physician would begin with a less t~an effective amo4~t for treatment and increase the dose until the desired effect is obtained exercising care to administer an amount less than the amount toxic to t~e host of the disease.
Both the above phar~aceutical composition and metnod of treat~ent include as active ingredient 9-(2-furfuryl)guanine.
The present inventicn also includes the novel intermediates as lollows:
(1) A compound of ~ormula III wherein R6 is alKyl of one to rour car~on atoms, aryl, or arylal~yl, n is zero or one; m is zero, one, t-~o, or three, ~ith tne proviso that 1~ or n is at least one; ~4 and ~5 are each independently hydrogen, al~yl of one to four carbon atoms, aryl, arylal~yl, or cycloalkyl of three to six carbon atoms, hydroxyal~yl of one to four `` ' 12~

carbon atoms, aryl, arylalkyl, or cycloalkyl of three to six carbon atoms, hydroxyalkyl of one to four carbon atoms, and Ar is heteroaryl or heteroaryl substituted by alkyl of one to four carbon atoms, alkoxy of one to four carbon atoms or halogen;
(2) a compound of Formula II wherein R6, n, m, R4, Rs, and Ar are as defined above; a~d (3) a compound of Formula IV wherein n, m, R4, Rs, and Ar are as defined above.
lQ Additionally, the novel processes of the present invention are as follows:
(A) A novel process for the preparation of a compound of Formula I, wherein Rl is OH or SH, R2 is NHR wherein R is as defined above, R3 is hydrogen, hydroxyl, mercapto, bromine, or NHR where R
is hydrogen or COR6 wherein R6 is as defined above, and n, m, R4, Rs, and Ar are also as defined above which comprises heating a compound of the Formula III
wherein R6, n, m, R4, Rs, and Ar are as defined above to obtain the compound of Formula I wherein R
is OH and R3' is NHCOOR6 wherein R6 is as defined above, and if desired, converting sai~ compound to a compound where Rl is S or SH by methods analogous to those known in the art, and if further desired, where R3' is N~COOR6 converting the compound to a compound where R3 is hydrogen, or NHR where R is hydrogen or COR6 wherein R6 is as defined above also by known methods.
Particularly, the above orocess is for the pre-paration of 8-amino-9-[(2-thienyl)methyl]quanine.
(~) A process for the preparation of a compound of the formula Rl a ~ N ~-- T

- 12~9~38 EDZ-~ -6-wherein Rl, R2, R, R3, R4, Rs, m, n, and Ar are as defined above, with the proviso that R3 is not Br, and R3 is not NHR; which comprises reacting a compound of the formula O
. ~
Hf ~ IV
~ N~ NH
Ar--~cH2)m ~ R4) ~ n with formic acid and formamide at elevated tempera-tures, to obtain a compound of the formula O

21~N
R N
Ar----tCH2 ~ 5) n-and, if desired, converting by methods analogous to those known in the art the compound into a compound of Formula I wherein Rl is SH and/or R3 is hydroxyl, mercapto, or NHR wherein R is as defined above or a pharmaceutically acceptable acid addition or base sal' thereof.
(C) A process for the preparation of a compound of the formula o ~ Br R2 ~ N

Ar- ~CH

r ~ 9~9L38 EDz-i -7-which comprises treating a compound of the formula Ar----~C~2 ~ ~) n with i~-bromosuccinimide in an organic solvent, and, if desired, converting by methods ~nown in the S art the resulting compound into a pharmaceutically acceptable acid addition or base salt thereof.
(D) A process for the preparation of a compound of the Formula I wherein R3 is `.~R
wherein R is as defined above; whicA comprises reacting a compound of the formula ~ (ca2~4 J
Rs n ~herein Xl, R2, R4, ~5, n, m, and Ar is as defined above; witn hydrazine at elevated temperatures and, optionally, ;~ith ~aney nic~el in l_ an alcohol solvent, and, if desir~d, converting the resulting compound where R is hydro~en to a compound where R is C0~6 with an al~anoyl halide, aroyl halide, or arylal~anoyl halide in tne presence of an organic base, and, if desired, 2~ whers Rl is.O or 0~, conv~rting said compound , ~2~

EDZ-l -8-to a compound where Rl i5 S or SH by known methods, and, if further desired, converting the resulting compound by methods analogous to those known in the art to a pharmaceutically acceptable acid addition or base salt thereo.
(~) A novel process for the preparation of a compound of Formula III wherein R6, n, m, R4, Rs, and Ar are as defined above which comprises contacting a compound of the Formula II wherein R6, n, m, R4, Rs, and Ar are as defined above, with a coupling agent in the presence of a solvent to obtain the compound of Formula III.
The coupling agent of the process is preferably N,~'-dicyclohexylcarbodiimide. The preferred solvent is anhydrous dimethylformamide.
(F) A novel process for the preparation of a compound of Formula II wherein R6, n, m, R4, Rs, and Ar is as defined abové which comprises refluxing a compound of Formula IV wherein n, m, R4, Rs, and Ar are as defined above in anhydrous methanol wlth anhydrous HCl then basified and treated with lower alkoxy carbonyl isothiocyanate to Qbtain the compound of Formula II.
(G) A novel process for the preparation of a compound of Formula IV ~herein n, m, R4, Rs, and Ar are as defined above which comprises:
step (1) reacting 2-amino-6-chloro-4-pyrimidinol in methoxyethanol with arylalkylæmine in the presen^e of triethylamine;
step (2) then treating the product of step (1) with aqueous ~odium nitrite, step (3) reducin~ ~he product of step (2) with sodium dithionite in formamide and 90% formic acid to obtain the compound of Formula IV.

~2~

EDZ-l~ -9-Under certain circumstances it is necessary to protect either the N or O of intermediates in the above noted process with suitable protecting groups which are known. Introduction and removal of such suitable oxygen and nitrogen protecting groups are well known in the art of organic chemistry; see fo~
example, (1~ "Protective Groups in Organic Chemistry,"
J. F. W. McOmie, ed., (New York, 1973), pp 43ff, 95ff; (2) J. F. W. McOmie, Advances in Organic Chemistry, Vol. 3, 191-281 (1963); (3) R. A.
Borssonas, Advances in Organic ~lemistry, Vol. 3, 159-190 (1963), and (4) J. F. W. McCmie, Chem. &
Ind., 603 (1979).
Examples of suitable oxygen protecting groups are benzyl, t-butyldimethylsilyl, methyl, isopropyl, ethyl, tertiary butyl, ethoxyethyl, and the like.
Protection of an N-H containing moiety is necessary for some of the processes described herein for the preparation of compounds of this invention. Suitable nitrogen protecting groups are benzyl, triphenyl-methyl, trialkylsilyl, trichloroethylcarbamate, trichloroethoxycarbonyl, vinyloxycarbamate, and the lik-.
Under certain circumstances it is necessary to protect two different oxygens wi~h dissimilar protecting groups such that one can be selectively removed while leaving the other in place. The ~enzyl and t-butyldimethylsilyl groups are used in this way either is removal~l2 in the presence of the other, benzyl being removed by catalytic hydrogenolysis, and t-butyldimethylsilyl being removed by reaction with, for example, tetra-n butylammonium fluoride.
In the process described herein for the prepara-tion of compounds of this invention the requirements for protective groups are senerally well recognized by one skilled in the art of organic chemistry, and -. ' "., :' . .

.. . ~L~

EDZ-l -10-accordingly the use of appropriate protecting groups i5 necessarily implied by the processes of the charts herein, although not expressly illustrated.
The products of the reactions described herein are isolated by conventional means such as extraction, distillation, chromatography, and the like.
The salts of compounds of Formula I described above are prepared by reacting the appropriata base with a stoichometric equivalent of the acid compounds of Formula I to obtain pharmacologically acceptable salts thereof.
The compounds of this invention may also exist in hydrated or solvated forms.
The above novel processes beginning with (G) and proceeding through ~F), and (E), or to and including (A) to obtain the compound of Formula I wherein Rl is OH and R3 is ~HCOOR6 wherein R6 is as defined above may be conducted in a one pot reaction.
Further, the lower alkoxy carbonylisothiocyanate may itself be added in the novel process (F) above or prepared in situ by suspending potassium thiocyanate in acetonitrile and adding methyl chloroformate to the suqpension for a mixture which is heated at reflux in the presence of the basified hydrochloride salt of Formula I in the above process (F).

DET~ILED DESCRIPTION

The compounds of Formula I and intermediates of Formula II and IV or the present invention exist in tautomeric forms as purines or suanines as illustrated below. Both forms ara included as part o~ tne inven-tion and are indiscriminately described in t'ne specificaticn.

3~3 EDZ - ' -11-.

~1 Rl R3 ~R3 ~2N N ~ 2N~N
Ar ~CX2t~ ~) Ar ~C~I2)m ~< ) Guanine Purine - I

~ CHCNHCOOR6 "
HN ~ _ _ N~ CXCNHCOOR6 l ~ 1 11 }I2N~N/ ~NH H2NJ~N/~ NH
Ar ~C~2) m~ 4 ) A~cH2t~R4\

II II

o OEI
HNJ~N~CHO ~ ~IHCXO

2~\N~ ~ H2~N
Ar (CX2~ 5 ) n ~<R

IV IV

. .
~ .

:~2~ L3~
DZ-l -12-The term "alKyl of one to four carbon atoms"
means a straight or branched hydrocarbon chain up to four carbon atoms such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl or tertiarybutyl. "~ydroxyalkyl of one to four carbon atoms" means the same alkyl radic:al with a terminal hydroxyl group.
The term "aryl n in_ludes an unsubstituted or substituted aromàtic ring such as, phenyl or phenyl substituted by halogen, e.g., fluorine, chlorine, bromine, or iodine, alkyl of one to four carbon atoms, such as methyl or ethyl, hydroxy, alkoxy of one to four carbon atoms, such as methoxy or ethoxy, or trifluoromethyl.
The ter~n "arylal~yl~ means an aromatic ring attached to an al~yl chain of up to four carbon atoms, ~uch as unsubstituted or suDstituted phenylethyl or benzyl where the substituents on the aromatic ring ma~
be the same as defined above.
2~ The te-rm "heteroaryl" means five- or six-.nembered aromatic ring containing one or more heteroatoms, such as nitrogen, oxygen and sulfur. Preferred radicals are the 2- or 3-furanyl; 2- or 3-thienyl; the ~
or ~-pyridyl; or 2-, 4-, or 5-thiazolyl radicals.
Pharmaceutically acceptable acid addition salts are those derived from inorganic acids such as hydrochloricr sulfuric and the liXe, as well as organic acids such as nethanesulfonic, toluenesulfonic, tartaric acid, and the like. These salts may also be 3~ prepared by standard methods known in the art.
Pharnaceutically acceptable base salt3 are those derived from inorganic bases such as sodium hydroxide, potassium hydroxide or ammoniun hydroxide or organic bases such as ar~inine, ~-methyl glucamine, lysine and the like. These salts ~ay also be prepared oy standard Inethods kno~n in the art.

8~
- EDZ-l -13-A preferred embodiment of the present invention is a compound of Formula 1 wherein Rl is OH or SH;
R2 is hydrogen or N~2; R3 is hydrogen, bromine or N~2; n is zero or one; m is zero or one, where n or m must be one; R4 and Rs are each independently hydrogen, alkyl of 1-4 carbon atoms or hydroxyalkyl of one to four carbon atoms, and Ar is Z- or 3-furanyl, 2- or 3-thienyl, or 2-, 3- or 4-pyridyl, oir 2- or 3-furanyl, 2-, 4-, or_~-thiazolyl, 2- or 3-thienyl, or 2-, 3-, or 4-pyridyl substituted by alkyl of one to four carbon atoms, or a pharmaceutically acceptable acid addition base salt.
Another preferred embodiment of the present invention is a compound o~ Formula 1 wherein Rl is ~; R2 is NH2; R3 is hydrogen, bromine or N~2; n is 0 or 1; m is 0 or 1, where n or m mus~ be 1, and Ar is 2- or 3-furanyl, 2- or 3-thienyl, ~-, 4-, or 5-thiazolyl, or 2-, 3- or 4-pyridyl, or 2- or 3-furanyl, ~- or 3-thienyl, 2-, 4-, or 5-thiazolyl, or 2-~ 3-, or 4-pyridyl substituted by methyl or etnyl, or a pharmaceutically acceptable acid addition or base salt.
Particular embodiments of--the present invention include:
g-[(3-pyridyl)methyl]guanine;
9-(2-thenyl~guanine;
9-[(2-pyridyl)methyl]guanine;
9-[(5-ethyl-2-thienyl)methyl]guanine;
-8-bromo-9-(2-thienylmethyl)guanine;
8-bromo-9-(5-ethyl-2-thenyl)guanine;
8-bromo-9-(2-furfuryl)guanine, 8-bromo-9-[(3-pyridyl)methyljguanine;
8-amino-9-(5-ethyl-2-thenyl)guanine;
8-amino-9-(2-thienylmethyl)guanine, and 8-amino-[9-(3-pyridyl)methyl]guanine.

"` ' 8~13~
EDZ-l -14-The most preferred compound is 8-amino-9-(2-thienylmethyl)guanine.
The 8-bromo compounds are not only useful pharma-cologically but are also useful as intermediates for preparing certain compounds of the present invention.
The compounds of Formula I may be prepared according to l~ethods B, A, and/or C as shown in the following Schemes 1, 2, and 3, respectively.
Generally, Method A is preferred.

3~

I. Scheme 1 - ~lethod B

HNJ13[No2 1~t ) Or~t n-e ~ o H NJ~N Cl ~C~2~10 ~ N2tl N NH, 2 Ar As O ' ,0 R~IL3~ b iSrtO 2 ~ H~
E~2NJ~N Cl H2tl H tt pa2520J
6 ~n~)n ~Drr~i~ acld ~CFt21 ID
Ar O O
tt ~o~nic dcid HNJ~3[NHC~lO
N ~o~amio~R Et 2N N NN

)n ~2) ~ tc~t2 Ar Ar ¦N aS /ACOH

HN~C ~>_8r _ H~ 3C ~ H ~r~ H~ N H2 t2N IN 2 H2 Ç~)n S~lR~)n S~ S~5 I CH 2 ~ H 2 ~ H 2 )~
r I ~ ~ ~ c no~-n ar:

H 9N~ NHCON~

n r ;ct ~.2~

EDZ~
II. Method a-~iscussion Compounds of Formula 8 above may also be used as starting materials and may be prepared by reacting 2-amino-6-chloro-4-hydroxy-5-nitropyrimidine, the compound of formula 2 described in J. Chem. ~oc., 1962, p. 4186, with the appropriate heteroaryl (al~yl) amine of formula 3 in tne presence of an organic ~ase at elevated temperatures. The resulting compound of the formula 4 is then treated with sodium dithionite -and formic acid followed by further treatment with formic acid and formamide at elevated temperatures to afford the compound of Formula 8.
Alternative, starting materials of Formula 8 may be prepared according to a modified method of C. ~. ~oelland, X. K. Robins in J. ;~ed. ~hem., 5, 55~, (1962) starting with a compound of the Formula 5 whicA
is reacted with an ap~ropriate heteroaryl alkyl amine of Formula 3, then with nitr~us acid to or.
the 5-nitrosopyrimidine, 6, which is reduced and rin~
closed by treatment with sodium dithionite, formic acid and formamide as described above.
The heteroaryl (alkyl) amines of Formula 3 are either commercially available or may be prepared by known methods.
Treatnent of a compound of Formula 8 with ~-bromosuccinimide in acetic acid, dimethyl~ormamide or ~ethanol ~roduces a compound of Formula la wAlcn when treated with hydrazine hydrate gives the hydrazine or directly the 8-amino derivative of Fornula lb. The reaction of the 8-bromo compound with hydrazine may or ~ay not proceed entirely to the 8-amino compound ~hus when the 8-hydrazine compound is obtained, it may be fur~her reacted with ~aney nicKel to allow tne red~lction to go to completion and . .

~L~2~

EDZ-~ -17-afford the desired 8-amino compound. Compounds of formula _ may be further converted by known methods to provide R6 substituents of formula ld ar, where Rl is 0, converting said compound to a compound of formula lc where Rl is S by reacting the said compound with P2Ss in presence of a base such a~
pyridine (examples given).

.: ~. , :

~z~

III. Scheme 2 - Method A
~F 'L"2 NH~3~ ~22)n tEtlN R~
82N N CL CH~O~OH, ~ tl25 C) 112N N Nll r~ f lux; 1 8h ~ ~r ---~CN 2~R ,~
IINO~ ¦ H256 2~ 2 ~ SJ n . Aq N~ O r. t .
A~OH ~-5h; 2 CN30N¦ ~R~) N-~520~;, HCOOH, 2 R n ¦bo~110~ 100-140 1 HCOOH H2NJ~N
J~r~CHl~ ~)n 4 ~R~)n _"_.. , .. , ~
CH3oNlHcl ~H) ~902 O s) IV~ NNJ~r 2-2HC~ NJkNN2 112N~NH '~q N2r~NN2 112Nl N hll 2+~< 5)n 6 ~RS)n ' ~Jhorr 51~-lt lif~
¦ R600C:ICS
O--tHNC0046 t tRf, -- C~3 or Zt) III H H~ t DCC ~NI i iHCOOli ~ C~ R5)n (n~d n~t l~ol~t-~ (n-~d noC t-ol~t~) "~ ~N ~--OH/IHCl IIN~N
pllcoo1lo ~ I I \~112~11 N2)~N~H ~ llth~ llzl~N~
~r ICHl~R45)n ._ . R5)r~

. ., 9L2~3~3 ~DZ--1 --19--IV. Scheme 3 - Method C

Cl Ar Br N NJ~

~ , N~ H2N~
Ar~CH 2~RS)n Ar~C~ 2~R5) n 1) Chromato- ¦ 2) HCl graph lc separaeion ~ ~ .

O O O
HNJ~ ~\ N~S HN~ ~ AqNH2N112 HNJ~
¦ I ~> ~ l l l ~ B r -~ H 2 H2N~N--~N~ H N~N~N~ R N~N~Nf Ar- ~C~2~ ) Ar (CE~2~ 4) Ar~cN2+~Rs) n 6 ~ 8 ~L~ 8S33L3~
EDZ-l -20-GeneraLly, the processes of the present inven-tion as shown in Scheme 2 above are as follows.
A 2-amino-6-chloro-4-pyrimidinol, that may be in the monohydrate form, is suspended in methoxy-ethanol, in the presence of excess amine or an organicbase such as triethylamine. The compound of Formula XXX having n, m, R4, Rs, and Ar as defined above is added to the suspension and heated optimally to a temperature at which the suspension refluxes.
Refluxing is continued until thin layer chromatography, for example, with 20% methanol in methylene chloride, shows the reaction producing a compound of Formula XX wherein n, m, R4, Rs, and Ar are as dsfined above i9 complete.
The reaction mixture naving the compound of Formula XX is then diluted with water and treated with sodium nitrite in the presence of acetic acid. A
nitroso of the Formula X again having n, m, R4, Rs, and Ar as defined above is obtained from the treatment by the nitrite as evidenced by a color change and precipitate. The temp~rature of the treatment is at about room temperature.
The treatment mixture having therein the nitroso of Formula X i5 contacted with sodium dithionite in a solvent mixture such as formamide, formic acid, at a temperature of from 6C-90C, preferably from 70-80C. m e tempera~ure is then raised to the boiling point of the solvent mixture, approximately 130-140C
for up to an hour, ~referably at least 20 minutes, or when the color of the nitroso containing mixture described above disappears and an inorganic salt precipitates. The product of this contact is ~
compound or t~e Formula IV wherein n, m, R4, R5, and Ar are as defined above.

3'`~ -EDZ-l ~21-Subsequently, the compound of Formula IV is dried and suspended in an anhydrous solvent such as methanol, ethanol, and the like. The suspension is then treated with a dry acid such as HCl, to form the acid salt shown as Formula IVa, wherein n, m, R4, Rs, and Ar are as defined above, or ~alt corresponding to the acid used for the treatment.
The salt IVa is basified with a concentrated mixture of NH40H~and 97% hydrazine to obtain a base of the Formula IVb wherein n, m, R4, Rs, and Ar are a~ defined above. The base is unstable, how~ever, i9 dried, for example in a vacuum over P20s.
The dried free base IVb is added to a solution of R600C~CS wherein R6 is as defined above. The solution of R600CNCS may be prepared by suspending potassium thiocyanate in a solvent suçh as acetonitrile and treating with slightly less than an equivalent of ClCOOR6 wherein R6 is as defined above at reflux for about one hour, cooled, then stirred to insure all of the alkylchloroformate is reacted before the base IVb is added. The reaction of dried free base IVb with the R6OOCNCS is mQ~itored to completion with thin-layer chromatography using silica in 20% methanol in methylene chloride to obtain a compound of Formula II wherein R6, n, m, R4, Rs, and Ar is as defined above.
A mixture of the compound or For~ula II and a coupling agent such as N,N'-dicyclohexylcarbodiimide, in a solvent such as anhvd~ous dimethylformamide, is stirred at about room t~mperature until completion of t~e reaction is shown by thin layer chromatograph to yield a compound of Formula III wherein R6, n, m, R4, Rs, and Ar are as defined above.

39~L3~

FDZ-l -22-The reaction mixture having the compound of Formula III and anhydrous potassium carbonate are suspended in a solvent such as anhydrous methanol, and refluxed until thin layer chromatography shows the reaction producing a compound of Formula I wherein R
is O or O~, R3' is NHCOOR6, wherein R6 is as defined above and n, m, R4, Rs, and Ar are as defined above.
The compound of Formula I wherein Rl, is O
and OH and R3' is NHCOOR6 may then, if desired be used to produce by known methods a compound of Formula I wherein R3 is NHR wherein R is as defined above other than CR6-Likewise, a compound of Formula I wherein R
is S or SH may be prepared by known methods fromthe compounds of Formula I wherein Rl is O or OH.
The preparation of compounds IV, II, III, and I
may be carried out in one pot. dowever, separation and purification of each of the compounds IV, II, III, or I may be effected by conventional methods, if desired.
Generally the processes of the present inven-tion as shown in Scheme 3 - Method C above ara as follows:
A mixture of 2-amino-6-chloropurine, potassium carbonate, and the starting material of the formula shown as 2 in Scheme 3 - Method C, that is generally commercially available or can be pre~ared by methods analogous to those known in the art, are stirred under nitrogen for from about 2 to 48 hours. A mixture of 7- and 9- substituted chloropurines shown as Formula 4 and Formula 5 in Scheme 3 - Method C are obtained.
The desired compound of Formula 4 is separated and treated with an aqueous acid such as dCl followed by addition of a weak solution of a base such as ~aOH.
The mixture is heated to assure it is neutralized ~ 28~
- EDZ-l -23-followed by conventional separation of the desired product of Fonmula I wherein R3 is hydrogen.
Subsequently, reactions to produce compo~mds of Formuls 7 and Formula 8 as shown in Scheme 3 -Method C are described above for corresponding stepsin Scheme I - Method B.
The compounds of the present invention have been ~hown to exhibit significant enzyme inhibition activity and cytotoxic activity. In the purine nucleoside phosphorylase (P~P-4) enzyme assay, total inhibition was achieved at a concentration less than about 300 micromoles on certain compound~ of the present invention. P~P-4 activity was measured radiochemically by measuring the formation of ~14-C]-hypoxanthine from ~14-C~inosine CBiomedicine, 33, 39 (1980)] using human erythrocyte as the enzyme source. The same compounds also were found by a standard test (HTBA-l) CScience, 214, 1137, (1981)] to be selectively cytotoxic for T-cells in the presence of 2'-deoxyguanosine at a sLmilar concentration range and nontoxic to B-cell in the presence of the same amount of 2'-deoxyguanosine.
Representative examples are shown in the activity ta~le.

.

.. ~ ~ , . .
, ~2~;3`~

EDZ-l -24-.

Activity Table Example Arl ¦ Method ¦ PNP-4 ¦ T-Cell ~dGuo ~umber ¦ LpreparationlICsO(~M)I(10 ~M) IC50 (~M
1 ¦3-PY ¦ B ¦ 21.9 ¦ 54.1 2 or 9a¦2-Th ¦ A or B ¦ 0.17 ¦0.83 3 ¦2-Th-5-Et ¦ B ¦ 0.93 ¦4.15 9b ¦2-FU ¦ B or A ¦ 0.25 ¦2.57 9c ¦3-Th ' ¦ A, B, or C¦ 0.085 ¦ 0.49 10 9d ¦2-Th-3-CH3¦ ~ ¦ 4.05 ¦ 8.6 16Ad ¦3-TH-2-CH3¦ A ¦ 1.72 ¦ 18.2 16Ac ¦CH2-2-Th ¦ A ¦ 6.25 ¦ 17.6 16Ak ¦3-TH-5-CH3l A ¦ 0.63 ¦ 2.8 16As l l A ¦ 8.45 ¦ ~12~5 1~ 1 1 1 15 9e ¦ ~ ¦ B ¦140 _ 16Ae ¦2-Th-5-Me ¦ A
16Af ¦2-Py ¦ A ¦ 4.6 lpy = pyridine, Th = ~iophPne, Fu = furan EDZ-l- -25-Since T-cells play a central role in immune response, use of the compounds of the invention is contemplated for the immunoregulation of autoimmune disease such as rheumatoid arthritis, systemic lupus erythematosus, inflammatory bowel disease, multiple sclerosis, myasthemia gravis, transplantation, juvenile diabetes, cancer, and viral diseases. The present invention thus includes composi~ions containing a compound of Formula I in treating disease such as autoimmune disease characterized by abnormal immune response in warmblooded animals. According to this aspect of the invention, the properties of the compounds of the invention are utilized by administering to a warmblooded animal an effective amount of a pharmaceutical composition containing as the active ingredient at least about 0.1 percent by weight, based on the total weight of the composition of at ieast one such compound of the invention.
Pharmaceutical compositions of the invention can be formulated in any suitable way, preferably with an inert carrier for administration orally, parenterally, ophthalmically, topically, or by suppository.
For example, the compounds o the present invention are formulated into dosage forms such as tablets or syrups by blending with an inert pharma-ceutical carrier such as lactose or simple syrup by methods well known in the art. For injec~able dosage forms, they are formulated with vehicles such as water, propylene glycol, peanut oil, sesame oil, and the like. I~ these dosage forms? the active ingredient is from about 0.05 grams to 0.5 grams per dosage unit.
The present invention is further illustrated by way of the ollowing examples.

, ' ~
':

3~1 EDZ-l -26-9-[(3-Pyridinyl)methyl]guanine 3-Pyridylmethylamine (15.8 ml; 0.1517 mole~ was added to a suspension of 2-amino-6-chloro-4-hydroxy-5 5-nitropyrimidine (14.45 g; 0.0758 mol) in isopro panol (600 ml). The mixture was heated under reflux for two hours and then stirred overnight at room temperature when the product, 2-am~no-4-hydroxy-6-[(3-pyridyl)methylamino]-5-nitropyrimidine, crystallized out~ The product was fil~ered, washed with water, and air dried.
The crude nitropyrimidine (25.32 g) from above was suspended in formamide (150 ml) and 90% formic acid (5~ ml) and the suspension was warmed to 70C
in a water bath. Sodium dithionite was carefully added to the ~arm suspension and then boiled for 15-2U minutesO The reaction mixture was diluted with hot water (300 ml), treated with charcoal and then boiled for an additional 20-25 minutes, filtered through celite, cooled and concentrated under reduced pressure to give formamido-~yrimidine which was collected by filtration, washed with- acetone, and dried under vacuum at 56C.
The above product was resuspended in formamide (100 ml) and formic acid (8 ml) and was heated under rerlux for 3.5 hours, poured onto 4~ ml ice-~ater and then filtered. Two crystallization from boiling water gave the analytical samyle of the desired product (4.5 g) mp >3uOC.

3~ EX~PLE 2 The procedure described in Example 1 was repeated to prepare the following 9-(heteroaryl or substitu~ed ~2~
EDZ-l -27-heteroaryl)methyl guanines, starting from appropriate hetaroaryl or substituted heteroaryl methylamines~
9-(2-Thienylmethyl)guanine, mp >300~.
9-[~2-Pyridinyl)methyl~guanine, mp >3~0C.
9-(~-Furanylmethyl)guanine, mp 296-299C, dec.
(~nown Compound: J. Am. Chem. ~oc., 1959, ~1:3U46.) 9-[(3-methyl-2-thienyl)methyl]guanine, mp >290C (dec).
9-[(2-methyl-3-~hienyl)methyl]guanine, mp >270~ (dec).
9-[(benzo[blthie`n-3-yl)methyl]guaIline~ mp >3~0C (dec).
9-(3-thienylmethyl)guanine, mp 320-3~2C (dec).

2-Amino-9-[(2-thienyl)meth~l]-~-chloro~urine A mixture of 2-amino-6-chloropurine ~Aldrich Chemical Co.) (7.~7 g; 0.44 mol), potassium car~onate (6.64 g; 0.~48 mol), and 3-thenylbromide (see ~S Patent number 3,74b,72~) (7.~ g; 0.~4~ mol) in ~F (200 ml) was stirred under nitrogen at room temperature for 4~ hours. The mixture was filtered and the filtrate evaporated to dryness under vacuum, et~yl ether was added and t~e precip1tate was collected by filtration to give a mixture oE 7- and 9-suDstituted chloropurines. A sample of ~ure 9isomer ~as prepared by chromatogra~hy on silica gel ~ith 5~
methanol/me~nylene chloride as the eluting solvent to separate it from the 7-isomer. Analytical sample -~as obtained by crystallization froIn acetonemethanol-mixture, yield 2.36 g, mp softens at 185C (dec) and then melts at 203-204C (dec).

~X~IYLE 2~

rhe procedure descri~ed in 2xample 'A was repeated to ;~re~ara the following 2-~mi~o-9-~(hetero-aryl or su~stitu.ei heteroaryl)metnylJ-6-cnloro~urines, ~2893L~3~
~DZ-l -2~-startiong from appro~riate heteroaryl or substit~ted heteroaryl ~ethylhalides.
2-Amino-9-[(;2,5-dimet~yl-3-thienyl)metn~11-6-chloro~urine, mp 190-192C (starting material 2,5-dimethyl-3-thenyl chloride was prepared according to the lit. proce~ure; ~uu-Hoi and Nguyen-~oan, ~ec.
Trav. ~him, 194~, 68:5).
2-Amino-9-(3-~uran~lmetnyl)-~-chLoropurine (starting material 3-furfurylchloride was prepared according to lit. procedure; S. P. Tanis, Tet. Letts., 1982, 23:3115).

EX~MPL~ 2C

9-[(2,5-dimethyl-3-thienyl)meth~l]~uanine A mixture of Z-amino-9-[(2,5-dimetnyl 3-thienyl)~ethyl]-6-chloropurine (3.8 g, ~.0129 mol) and 2N ~Cl ~as heated on a steambath for 3.0 hours and then heated under refl~x for another hour. At the end of this time 1~ i~aO~ solution was added to the solution till basic and the mixt~re was heated 2U for another five minutes. The mixture was then acidi~ied with acetic acid, cooled, and ~iltered to give 3.6 g, of the product. ~n analyticai sample was obtained by chromatography over silica gel using 10 metAanol/chloroform as eluting solvent, mp >3UuC
(dec).

~XAi~PL~ 2D

The procedure descriked in ~xample 2C was re2eated to orepared 9-(3-tnenyl)guanine, m~ 3Z0-322~ (dec).

.

E ,YA.~IP LE 2 r' 9-(3-furanylmethyl)~uanine The crude 2-amino-9-~3-furfuryl)-6-chloropurine (4.74 g, 0.019 mol) was suspended in methanol (175 ml) and a solution of sodiu.~ methoxide, prepared from sodium metal (1.75 g; 0.07~ g atom), and methanol (75 ml), was slowly added to the suspension, followed by 2-mercaptoethanol (6.1 ml = 6.8 g; 0.0~7 mol) and water (0.35 ~1). The reaction mixture was heated lU under reflux (N2 atm) for two hours, when an additional amount of sodium methoxide from 1.14 g sodium (0.05 g atom) and 25 ml methanol was added.
After an additional 2~5 hours reflux, the reac~ion mixture was concentrated under vacuum to 75 ml and then diluted with water (20~ ml), and acidified with acetic acid (pd 5.5). The white ppt. was filtered, washed with water, and dried, yield 4.05 g; mp 308-310~.

EX~ LE 3 9-~(5-Ethyl-2-thienyl)methyl~quanine 2-~mino-6-chloro-4-~yrimidinol, monohidrate (2~.96 g; 0.1193 mole) was suspended in methoxyethanol (3~0 ml) and 5-ethyl-2-thenylamine (16.5~ 9;
O.li53 mole) prepdred from 2-ethyltnio~nene accocdlng to the Lit. Procedure, (J~S, 1948, 70:4018) t~as added - to the suspension. The resulting solution was heated under reflux for one hour and tAen 16.~ ml of triethylamine was added and the refluxing continued for an additional 18 noura~ The reaction mixture was ~oured into ice ~ater (bOO ml), diluted wit:~ acetic acid (100 ml) and then tredted with a solution of ., .

EDZ-l ~30-sodium nitrite (16 g) in ~ater (1~ ml). The mixture was stlrred at room temperature for 1.5 hours and the resulting salmon colored nitroso compound was collected by filtration and washed wit~ water.
The crude nitrosopyriJnidine was then reduced with sodium dithionite in formamide (200 ml) and 90~ formic acid (1~0 ml) at 70C and then boiled for 20 minutes. The reaction mixture was diluted with water ~300 ml) and the boiling continued for an additional 30 minutes, filtered hot, and then allowed to crystallize in the refrigerator. The crude N-formyl derivative (28 g) was collected by filtration, washed with ~ater and air dried and then cyclized witb formic acid (1~ ml) and formamide (10~ ml) at reflux te~perature for four hours. The hot reaction mixture was ~oured into 500 ml of ice water to give t:~e cr~de quanine which was then purified by dissolving in boiling 1.5 ~ treatin~
with charcoal and thesl precipitatin~ with alnmonium hydroxide. rhe crude yuanine was t:~en redissolved in hot 1 ~ .~a~ solution, treated witn cnarcoal, filtered and the filtrate acidified witn acetic acid to give the desired product ~hich was used in tAe next step without ~urther purification.

EX~PL~ ~

The procedure descrioed in ~xample 3 ~as re2ea~ed to prepare 9-~2-thenyl)guanine, m~ >30~~ starting from 2-al~ino-6-chloro-~-pyrimidinol and 2-thenyla~ine.

~23~ 3~
~DZ-l -31-2X~PLE S

8-Bromo-9-(2-thienylmethyl)quanine N-3romosuccinimide (2.~2 g; 1~.7 mmol) was added to a cold (0C) suspension of 9-~2-thenyl) guanine (3,5 g; 14.1 mmol) in DMF (1~0 ml) and the mixture was stirred ~or 30 minutes at 0C
and then at room temperature for 24 hours. The reaction mixture was then diluted witn 75 ml of water and filtered. Recrystalllzation of the lU product from D~F gave the analytical sample, yield 3.1 g; mp ~9~-295C (dec).

EX~MPLE 6 The procedure described in 2xample 5 was re~eated to prepare the following 8-~romo-9-[(substituted heteroaryl)methyl]guanines, starting from appropriate 9-L(substituted heteroaryl)methyl]guanines in each case.
8-oromo-9-~5-e hyl-2-thienylmethyl)guanine 8-bromo-9-(2-furanylmethyl)guanine, mp >3~0C.
8-bromo-9-[(2-methyl-3-thienyl)methyl]guanine, mp >280C (dec).
3-bromo-9-~(o2nzo[blthien-3-yl)methyl]guanine, mp 25~-2~0C (dec).

~XA.~PL2 7 8-Bromo-9-[(3-pyridin~)methyl]guanlne N-Bromosuccinimide (1.59 g; 8.95 ~mol) was addea to a suspension of 9-[(3-pyridyl)~ethyl~guanii~e (2.0 g; 8.14 i~mol) in glacial acetic acid (20 ml) ~n~
the mixture was stirred at room emperature LOr 3U 3.5 Aours. The reaction ~ ture ~as concentrated , ~2~L3~1 -EDZ-l -32-under reduced pressure and then diluted with water and fil~ered. The crude product was triturated with water, filtered, and washed with water and dried.
Yield 1.91 g, mp ~300C.

EX~PLE 8 .
8-Amino-9-(5-ethyl-2-thien~lmeth l)guanine A mixture o~ 8-bromo-9-(5-ethyl-2-~enyl)guanine (6.5 g; 18.3 mmol) and 60% aqueous hydrazine ~200 ml) was h~ated to reflux under nitrogen atmosphere for 20 hours. 2-Methoxyethanol (50 Ml) was added and the refluxing continued for an additional 48 hour~ in the open air. The orange-brown solution was cooled, diluted with water (150 ml) and allowed to crystallize in the refrigerator overnight. The crude product thus obtained was converted to the hydrochloride salt by recrystallizing from boiling isopropanol and 1 N HCl.
Yield, 0.49 g; mp 215-218C, dec.

~MPLE 9 The procedure described in E~ample 8 was repeated to prepare the followiny 8-amino-9-[(substituted heteroaryl)methyl)guanines, starting ~rom appropriate 8-bromo-9-~(substituted heteroaryl)methyl]guanines:
8-Amino-9-~(3-pyridyl)methyl]guanine, mp >300C and additionally, the following compounds a through e were prepared.
a. 8-Amino-9-(2-thenyl)guanine or 8-aminc-9-[(2-thienyl)methyl]guanine as hydrochloride sal', 0 5 ~2~ mp 223-226C (dec).
b. 8-Amino-9-(2-furanylmethyl)guanine monohydro-chloride, 1.0 H20, m~ 197-19~C (dec).
c. 8-Amino-9-[(3-thienyl)metnyl]suanine, monohydro-chloride, monohydrate, mp 275-278C ~dec).

~28~

rDZ~~ -3~-d. ~-Amino-9-[(3-methyl-2-thienyl)methyl]guanine, 0.25 ~O, mp 290C (dec).
e. 8-Amino-9-[(benzo [D] thien-3-~l)methyl]guanine, 0.5 H2O, mp >300C (dec).
Starting materials, such as 2-, 3~, or 4-pyridylmethylamines, 2-thenylamine also called 2-(aminomethyl)thiophene or ~-thiophene methylamine, and 2-furfurylamine are com;nercially availa~le (for _ example, Aldrich ~hemical Company). The suDstituted thenylamines were synthesized from the su~stituted thiophenes using a general literature procedure.
(~. D. ~artough and S. L. Meisel, J. Am. ~hem. Soc., 19~8, 70:4018).
2-Amino-6-chloro-4-hydroxy-5-nitroQyrimidine lS was synthesized according to a method described in the literature (A. Stuart and H. C. ~. ~ood, J. Chem. ~oc., 1963:11~6), 2-~nino-6-chloro-4-~yrimidinol monohydrate was purchased from Aldrich Chemical ~ompany.

~ PLE l~

2-A~ino-4[~(2-thienyl)meth~l]amino~ (formamido)-6-~yrimidinol ta compound of ~cheme 2 Formula I'J
wherein n is one, m is zero, R~ and R~ are hvdroqen, Ar is ~-thien~
2-~nino-6-chloro-4-pyrimidinol, monohydrate (85~, 100.0 g, 0.5191 mole) was suspended in metho~yethanol (700 ml) and 2-thiophenemetAylamine (9~, 61.3 g, 0.~1~7 mole) is added to the suspension. The mi~ture was heated under reflux for two hours and then 73 ml 3~ (d = 0.725, ~.52 mole) of triethylamine ~as added and the refluxing continued for an additional 13 hours.
(The reaction was followed by TL~: 20~ metnanol-C~13.) The reation Inix_ure was roured lnto ice water (1~00 ml), diluted with acetic acid (~00 ml), and tnen - ~8~

EDZ-l -34-treated with a solution of sodium nitrite (80 g, 1.16 mole~ in water (300 ml). The mixture was stirred at room temperature for four hours, and the resulting reddish colored nitroso compound (X) was collected by filtration and washed ~ith water (the reaction was followed by observing the color change in the forma-tion of the precipitate).
The crude nitrosopyrimidine of Formula X, (of Scheme 2, Formula X, wherein n is one, m is zero, R4 and Rs are each hydrogen and Ar is 2-thienyl) prepared above was divided into two batches and each in turn was then reduced with sodium dithionite (>90~
70 g, 0.36 mole) in formamide (300 ml) and 90~ formic acid (300 ml) at aooc and then boiled for 20 minutes.
The temperature was approximately 130-140C at this point. The reaction was complete when the red color completely disaQpears and inorganic salt precipitates.
The reaction mixture was diluted with water (300 ml) and the boiling continued for an additional 30 minutes, filtered hot, and then allowed to crystallize in the refrigerator. The reaction was monitored to completion by TLC (sio2; 20~ C~3O~ in C~C13. The crude ~-formyl derivative, 2-amino-4~(2-thienyl)methyl]amino~-5-(formamido)-6-pyrimidinol, (100 g) was collected by filtration,washed with water, and dried and used in ~he next step without further purification in most cases.

E~MPLE lOA

~he procedure dessribed in ~xample 10 was repeated to Qrepare 2 ~mino-[~(3-thienyl)-methyl~amino]-5-~formamido)-6-pyrimidinol starting from 3-thiophene methylamine and 2-amino-6-chloro-4-~yrimidinol.

~. ; .
.:.

- ~L2~
EDZ-l ~35~
E.YAL~PLE 11 ~-Amino-4-[[~2-furanvlmethvl)amino]-5-(formamido)-6-pvrimïdinol A mixture of 2-amino-6-chloro-5-ni~ro-4-pyrilni-dinol (J. CHem. ~oc., 1962, p 4186) (31.5 9;
0.15 mol) methanol (1~00 ml) and Eurfurylamine (2~.1 g; u.3 mol) was stirred and heated under reflux (i~ atm) for six hours. The reaction mixture was cooled, filtered, washed with water, and air dried to give 34.43 g of yellow solid, mp 286-2~gC
(dec), which was used in the next reaction.
The crude nitropyrimidlne (33.9 g; 0.135 mol) was suspended in formamide (2g~ ml) and 88-~ formic acid (145 ml), and then warmed up to 80~. Sodlum ditAionite (~7 g; 0.327 mol) was slo~ly added to the war~ (8b-850C) susoension over a period of 50 ~inutes, maintained at the tem~erature (~ 85C) lor another 3~ minutes, then diluted with boiling water (12~ ml), and heated the mixture around 850r for ano~her ~u minutes when tan colored crystals were ~ormed.
The product was filtered off, washed ~it~ ~ater, and dried over P20s under vacuum ~vernight. ~ield, 23.~ g, mp 24~-247C (dec). In most cases tnese compounds ~ere carried through the reaction se~uences 2~ witAout characterization.

E~2LL~IPL~ 11~j ~ he procedure descriL~ed ir. ~xample 11 was repeated to pcepare the ~ollowing 2-amino-4Ll(heter aryl or substituted heteroaryl)methyl]amino]-~-3u (f~rmamido)-6-pyrimidinols, (~'a~ie 1) starting Crom appropriate heteroaryl or substituted neteroaryl methylamines and ~-amino-6-chloro-~-nitro-4-pyrimidinol.

- ~21!~

EDZ-l -36-T~L~ 1 NHCHO
~N / ~
N--~N~H
Ar Ar or Ar 2-Thienyl S

3-Thienyl '' ' ~,3 "' 3-Furanyl ~9 1~ (2-thienyl)methyl ,~

3-;~e-2-thienyl ~ e-3-t~ienyl ,~3 , .D~.-l ~37~
TA3~E 1 (C3NT'~) ~r or Ar 5-1~le-2-thienyl ~ S CH3 5 2-Pyri~ inyl Bel~zoLb~thien-2-yl 2-tniazolyl 1~ _ N
/(~S~

4-thiazolyl N/

(~-thienyl)methyl H2C ~

15 5-Me-3-thienyl Ps ~ c~3 12~
EDZ-l -38-TA~LE 1 (CO~

Ar or Ar 5-l~e-2-furanyl C~3 4-i~e-3-thienyl CX3 S

4-~e-2-thienyl CH3 /~S

EX~MPL~ 12 2,5-Diamlno-4-[(2-thienylmeth~l)amin~]Qyrimidin~6-ol, dih~drochloride (a com~ound of ~chelne 2, r~?nnula I~la, wherein n is one, ~ is zero, R4 and Rs are edch hydrogen, and ~r is 2-thienyl) ~he crude N-formyl derivative as prepare~3 in Fxam~le 1~ aDove (40 g, D.1508 .-nole) ~as ~uspended in anhydrous methanol (500 ml) and a stream of dry ~Cl (g) was passed tnrough the solution wnile r.ea~ing th- mixture at reflux. The reaction was continued foc 2.5 hour when a clear solution was forme~ fo110wed by 2u a crystalline precipitate. The mixt~re was cooled i!l all ice bath and then filtered to give tile salt, 2,5-diamino-4-[~2-thienyl.~e~nyl)amin~]~yriinidin-6-ol, dihydrochloride, (~.6 ~). Concentration of the mothec li~uor gave an addl~-onal amount of tne sal~

.
:-~DZ-l -39-(8.65 g). Total yield, 37.25 g (79~). The material was carried on without further purification.
Alternatively, the N-formyl derivative was refluxed with 5~ methanolic-~Cl (g) to give the desired diamine- 2 HCl salt.

EXA.~PLE 12A

The procedure described in ~xample l~ w~-~epeated to prepare the ~ollowing 2,5-diamino-4-L[(heteroaryl or substituted heteroaryl)methyl]amino~æyrimidin-6-ol, as dihydrochloride salt (Table ~), starting fromappropriate 2-amino-4- L L (heteroaryl or substituted heteroaryl)methyl]amino~-5-(formamido)-b-pyrimidinol (Table 1).
TABL~ 2 HN ~ 2 HCl ~2N ~ N NL
Ar Ar or ~r 3-Thienyl 2-Furanyl o 3-~uranyl ~3 , - lZ~L3~ -- ~DZ-l ~40-TA~LE 2 (CO~T'D) Ar or Ar (2-thienyl)methyl ~¢~

5 3-Me-2-thienyl H3C
~3 2-Me-3-thienyl ~\
H3C ~ S

5-Me-2-thienyl ~
~/~

2-Pyridinyl 1~N - )J

s2nzo[~]thien-2-yL
~5r 2-thid7olyl i~
~S~

. .
.
.~ ., : -, " ', ~ '' : ' .

~28~
- ~.DZ-l -41-TA~L~ 2 (CONT'~) Ar or Ar 4-thiazolyl N

S

(3-thienyl)~ethyl S-!~le-3-thienyl ~ ' 5-:~e-2-furanyl 4-~e-3-thienyl ~ H3 4-~e-2-thienyl ~3 S~

, ~, ~"' .

: "
:' EDZ-~ 4~
EX~PL~ 13 ~ethyl [[[2-Amino-1,6-dihYdro-6-oxo-~-[(2-thien~l-methyl)amino]-5-e-yrimidinylJamino]thioxome~h~1]-carDamate (A cor~lpound of Schene 2, Formula II
wherein Rh is meth~l, n is one, M iS zero, R4 and R~ are eacn hvdro~en and Ar is 2-thienyl~
The crude di;lydrochloride salt as prepared in ~xample 12 above ~37.2 g, ~ mole) was suspended in water (300 ml) and then basified witn a mixture l~ of concentrated N~40~ and 97% hydrazine ~3:1) (40 ml) to give the free base which was dried ovar vacuum over P20s for 20 hours. Yield 26.1 g (975i) of the base shown as compound of ~cheme II, Formula IV~ wherein n, m, X4, ~5, and Ar are as defined above. This free base was unstable.
A suspension of potassium thiocyana~e (l~
O.i~6 mole) in acetonitrile (25~ ml) was treated with methyl chlorofor~ate (99~i) (13.8 ml, 0,177 mole) and the mixture ~as heated at reflux for one Aour, cooled, and then filtered to remove inorgallic s21ts.
The oright yellow colored filtrate is~ 3tirred overni~ht at room te.n~erature under nitrogen. Care should oe taXen so that all of the methylchlorofor~ate has reacted '~efore proceeding. The dry base (26.1 ~) ~as added to the solution of methoxycarbonyl isothiocyanate and the stirring continued for 36 nours at room temperat~re under nitroyen. ~he reaction was moni-tored to completion hy TL~ (si~2; ~ C~3~ in CHCl3). The product was filtered of_ and -~asned 3~ ~ith ;nethanol to give th~ thiourea derivative, metAyl [[[~-amino-1,6-dihydro-6-oxo-4-[(?-tnienyl.nethvl)-amino]-5-pyrimidinyl~alnino]thioxomethyl]car3ainate.
Yield 37.4 g (96~), mp 225-2~C; (~ ure by HPLC).

- . ~28~3~

EDZ-l ~43 Alternatively, the nitro or nitrosopyrimidines were ca~alytically reduced and rea~ted i~nediately with ethoxycarbonyl isothiocyanate to give ~he thlo-ursa derivative.
This material was carried on to the next step without further purification.
.

E~AMPLE 13A
. ._ _ ~ . . .
The procedure describ~d in Exampla 1~ was repeated to prspare the fol}owing methyl ~or ethyl~
[LL2-anino-l~6-dihydro-6-oxo-4-cL(heteroaryl or 5 ubstituted heteroaryl)methyl~amino]-5-pyrimidinyl]-amino]thioxo-nethyl]car'oamate (Table 3) stArting from appropriate 2,5-diamino-4-C.(heteroaryl or substituted heteroaryl)methyl~anino]pyr~nidin-6-oL, dihydrochloride ~alt (Table 2).

. .

- ~28913~
EDZ -L ~44~

HN ~ NH--C-NHCOO R6 H2N~NJ~ NH
AY' Ar orAr R6 2-Thienyl ,~3 Et S

3-Thienyl ~3~ CH3 2-Furanyl ~3 CH3 -3-Furanyl ~ CH3 mp 246-249C
O ( dec) ~2 -thienyl ) methyl ~ C~I3 --H2C~l\S~ 234-239 C

2--~e- 3 -thi enyl ~ C~ 3 np 235-237C
C \S/ ( ~ec) -, ~
'". :

: ' :

EDZ~ 5~
TABLE 3 (CONT'D) Ar or Ar R6 5-Me-2-thienyl ~ CH3, S ~ CH mp 227 230C
(dec) 2-Me-2-thienyl " , Et 2-Pyridinyl ~ CH3 Benzo[b]thien-2-yl ~ CH3 2-thiazolyl ~ ~ CH3 S mp 211-215C

15 4-thiazolyl ~ ~ CH3 mp 217-218C

(3-thienyl)methyl ~ CH3 20 5-Me-3-thienyl ~ c~3 ~2~g~3~
EDZ-l ~4~
TABLE .~ (CONT'~) Ar or Ar R6 5-Me-2-furanyl CH3 ~ mp 228-229C
\ O ~ CH3 (dec) 4-Me-3-thienyl ~ CH3 C'~3, Et S

4-Me-2-t~ienyl ~ c~3 CH3, Et S

. EXAMPL~ 14 M~thyl ~5~amino-7 [(2-thienylme nyl)~nino]-oxazoloC5,4-dJ~yrimidi_-2-yl]carbamato (52e Scheme 2, Formula III wherein R~ is methyl, n is one, m is zero, R4 and Rs are each h~dro~en and Ar is 2-thieny~) A mixture of the thiourea derivative as pre~ared in Example 13 ~35 g; 0.096 mole) and N,N'-dicyclohexylcarbodiimide (DCC) (~9.4 g, 0.288 mole) was suspended in dry DMF (1800 ml) and stirred at room temperature for 24 hours. The course of tne reaction was followed by TLC (SiO2, 20~, C~3O~
in CHC13). The DMF was compls~ely strippe-~ off under vacu~m and the -esi~ue .-iturated twic~ with ., ~ . :

., .

: .

~2~9~3L3~3 ED~ 47-C~2C12 to give tne desirad carbamate, methyl [5-amino-7-[~2-thienylmethyl)amlno~oxazolo[5,4-d~-pyrimidin-2-yl~carbamate. Yield 27.~ g (90~), mp 00C. Purity 97.6~ (~PLC).
This material was carried on to the next step without further purification.

E~MPLE 14A

The procedure described in Example 14 ~as repeated to prepare the followin~ methyl (or ethyl) 1~ [5-amino-7-[~(heteroaryl or sub~tituted heteroaryl)-methyl]amino]oxazolo~5,4-d]pyrimidin-2-yl]carbamate (Table 4) st3rting from appropriate methyl (or ethyl) ~2-amino-1,6 dihydro-6-axo-4-[~(hateroaryl or substituted heteroaryl)methyl]amino]-5-pyrimidinyl]-lS amino]thioxomethyl~carbamate (Table 3).

- ~2~ 3~38 EDZ--l --4 8--~NI}COOR6 H 2NJ" L Ar Ar ~ or Ar R6 2-Thienyl ~3 Et S

3-Thienyl ~3 CH3 2-Faranyl CE13 3-F-~lranyl i CH3 mp 28~3-291 _ 0 (dec) 1~ .
( 2 -thi ~nyl ) methyl CH 3 //~ mp ~270C
--H C/~S / ( dec ) 23 2-Me-3-t;-1ienyl \~ C~3 ~/ ~ mp >270C
H 3C \S ( dec ) . ~

EDZ-l -49-TAB T .~ 4 (CONT'D) Ar or Ar 5-Me-2 thienyl CH3, Et ~ p ~250C
~ \ S~~~C~ (dsc) 2-Pyridinyl ~ C~3 BenzoLbjthien-2-yl ~ I ~ CH3 .

2-tniazolyl N ~ CH3 /~S~ -4-thiazolyl N ~ C'~3 ~ ~

(3-thienyl)methyl CH3 ---H2C~____ ~S~' 5-L~e-3-thi2nyl CH3 ~ S ~ C~3 ~L2~9~L3~

TABLE 4 (CONT'D) Ar or Ar R6 5-Me-2-furanyl CH3 ,~
/ \ O ~`CH

5 5-.~e-2-thi~nyl ~ C~3, ~t 5-~e-3-tnienyl ~ ,CH3 C~3 ~S~ ' 4~e-2-thienyl c~3 Et _ mp ~2~0C

E ~I~LE 15 ~ethyl [2-amino-6,9-dihvdro-6-oxo-9-(2-thieny1methyl-~ urin-8-yl]car~nate (See ~che~e 2, For~ula I
wherein R~ is NHCOOR6,~he~ Rs is ,neth~1, n is one, In is zero, R4 and R~ are each hydro~en an Ar is 2-thien~1) .
A mixturs of the oxa~olocarbamate as prepared in ~xalnple 14 (25 g, 0.078 mole) and anhydrous K2CO3 was su~pended in anhydrous ~ethanol and hea.3d to reflux for eiynt hours. 'Fne course or t'ne reac~ion 9~3~3 EDZ-l -51-is being followed by the TLC system mentioned above.The reaction mixture was then evaporated to dryness under reduced pressure and the residue dissolved in ammonium chloride solution (16.8 g; 0.312 mole in 200 ml of water). The resulting precipitate was collected and dried giving 24.:39 g of the metnyl[2-amino-6,9-dihydro-6-oxo-9-(2-thienylmethyl)-lH-pyrin-8-yl]carbamate, somet mes contaminated with the 8-amino compound, i.2., in this example, 89.58~
carbamate and 9.54~ 8-amino compound, 8-amino-9[(2-thienyl)methyl]guanine of Formula I wherein R3 is m is material was carried on to the next step without further purification.

The procedure described in Example 15 was repeated to prepare the following methyl (or ethyl) [2-amino-6,9-dihydro-6-oxo-9-[(heteroaryl or substituted heteroaryl)methyl-l~-purin-8-yl~-carbamate (Table 5) starting from appropriate methyl(or ethyl) t5-amino-7-~(heteroaryl or substituted heteroaryl)methyl]amino]oxazolo~5,4-d~pyrimidin-2-yl]-carbamate (Table 4).

~, ..

39~L38 .
EDZ-l -52-N
HcooR6 Ar Ar orAr ~

2-T~ienyl Et, ~np ~ >250C (dec) S

3-ThienyL C~3 2-~uranyl C~3 ' ~ ~ .np >300C
~ o / (dec~

3-~uranyl ~ CY.3 ~p >270C
0 (dec) (2-~ienyl)methyL c'~3 ~2C S

. 2-~'12-3-t.liesly~ C~13 20, ~ C~ \S~

. . , , ' ,:

' , ' ` ~' '' " '' : ' ' ' .

12~3~3L.3~

- ED'Z-l -53-TABLE 4 ( CONT ' ;) ) Ar or Ar ~6 5-Me-2-thienyl CH3, Et ~/~ illp >250 C
~\S/\C~3 (dec) 2-Pyridinyl 1~ CH3 N
3enzo~b]thien- -yl [~ CL{3 2-t~i~zolyl N~ CH3 S

4-th~ azolyl N--\~ C~3 l~S/

(3-~nie4yL)~e:hYL --r42C~3 c~3 5-M2-3-t~ienyl ~ Cli3 S C~I3 .3~3 - EDZ-l ~54~
TABL2 4 (CONT'~) Ar or Ar R6 5-Me-2-furanyl C~3 ~ mp >250C
/ \ O ~ C~3 (dec~

4-.~e-3-thienyl~ -CH3 CH3, Et S

4-Me-2-tnienyl~ CH3 C~3 Et Il /\S/

EXAMPLE 1~ -8-.~ino-9-[(2-t~ienyl ? .-nethyl]~uanine (See Sche,ne 2 Forinula I ~herein R~ is ~, n is one, l~ is zero, R4 and ~ are each hvdro~en and Ar i~ 2-~hienyl) The crude carbamata as ~reparsd in Exa,nple 15 (a9.5~ t'ne carbama~e ;~lus 9.5~ the ~-a~-n~no colnpound (20.39 ~; 0.064 nole) r~ the previous reac.ion is suspended in isopropdnol (125 ml) and 1 U ~Cl ~125 ml;
0.125 mol~) anil the mixture heatad at reflux for -20 hours (the reaction is ,nonltore~ '~y TLC (SiO2:20~
~e'~H in C~C13, CH3C~:~.OAc:H2O ~:l:l)j, wnen a clear solutio.n is for~e~. On _oolin~ the ?roduct crystallizss ou~ 'ro~n tna solution as the ... ..

EDZ-l ~55~
hydrochloride sal~ of 8-amino-9[(2-thienyL)-methyl~guanine. Yield 15.1 g (76~). Purity 98~ by ~PLC, mp 219-222~C (dec).
The hydrolysis was also c:arried out in 10~
methanolic sodiuln hydroxide solution under reflux tenperature, followed by neutralization and recrystal-lization from appropriate solvent.

. , .
The procedura described in Example lo wa~
repeated to prepare the following 8-amino-9L(hetero-aryl or substituted heteroaryl)methyl~guanines (Table 6) starting f-om appropriate methyl (or ethyl) [2-amino-o,9-dihydro-o-oxo-9-[(neteroaryl or substitlted heteroaryl).nethyl~ purin-8-yl~-carbamate (Table 5).

TABL~ 6 2 ~ `'H ~

Ar or X ,~pC

2-Fur~nyl ~ '~Cl 253-7 a. 3-~hianyl ~r--~ H~ 20 275-d S

3~

E~Z-l -;6-TA3L~ 6 (CONT'D) Ar or Ar X mpC

b. 3-Furanyl ~ H l 293 4 ~ ~ (d) S O

. . _ . . .

c.(2-thienyl)methyl ~Cl H20 153-5 (d) 10 d. 2-~e-3-t~ienyl ~ '.~Cl 26~-~

H3C ~ S

e. S-Me-2-t~ienyl -~Cl 0.25 ~260 -* ;,~0 lS ~ S C~3 f. 2-Pyridinyl ~ 1.5 -HCl 270-2 N ~ - 0.25 J2 (-i) ~. 3enzolojthien-2-yl 0.25 ;~2 ~300 ~ ~ (d) . ; :

TA~LE 6 ( CO~T ' ;~ ) Ar or Ar X ~pC

h. 2-thiaz~lyl N 1.2 HCl >250 ~3 1.2 ~2 S

i. 4-thiazolyl N ~ 1.1 HCl ~250 ~ ~ 0. 3 ~2 j . ( 3 -thienyl ) methyl 0. 9 EICl ~ 177-~3 H20 (d) ~2C~, k. 5-Me-3-t~ienyl HC1 0.5 H20 212-;
\ (d) l; ~
- S /~C~

L. ;-~e-2-furanyl ~Cl-1.15 212-~
~ ~2 (d) / ~ o CX3 12~ 3~3 EDZ-l -5~-T~3LE 6 (CONY"D) Ar or Ar X mp C

m. 4-Me-3-thienyl 0.~ HCl ~240 ~ C~3 1.25 H20 (d) n. 4-Me- ~ ienyl ~

E~IPL~ 17 2,~-Dianino-l,9-dihydro-9-(2-thienyl~neth~1-6H-~urine-6-t~ione A mixture of P25s (2.4 g: 10.95 mmol), pyridine (30 .~1) ~nd 8-amino-9~2-t;~ienyl)methyl~-guanine (1.5 g; 4.a7 mmol) was heated under reflux l; for 4.5 hours an~ then poured int~ 200 ml of boilin~
water and boilad for one nour. The mixture was allowed to stand at room t2mperature overnight. The precipitated solid ~as collected, diss31ved in 1 ~
NaOH, traated wl~h ac~ivated charcoal, filtered, and 2~ t;~en acidiried with ylacial acetic acid to pH 5.4.
ne precipitated solid was c~llected, ~is301ved in 1 ~ HCl, t~eated with ac_ivarad charcoal, filtered, and .~eutralized wit~ NH4~H to pH 7.~7 to giv2 5~2 m~ o_ the deslred product, ~p >300C.

- - 12~9~38 EDZ-l -59~
EX~PLE 17A

The procedure described in Example 17 was rapeated to prepare tne following 2,8-diamino-1,9-dihydro-9-[(heteroaryl or substituted heteroaryl)-methyl]-6H-purin-o-thione, starting from appropriata 8-amino-9[(heteroaryl or -ubstituted heteroaryl)alXyl]
guanine.
2,8-Di~ino-1,9-dihydro-9-(3-thienylmethyl)-6H-purine-6-thione, 0-5 H2O~ mp 275~ (dec).
2,8-Diamino-1,9-dihy~ro-9-~2-(2-thienyl~ethyl]-6a-purine~6-~ione, 0.25 H2O, mp >260C (dec).

EXAMPLE 1 a 2-~ino-?,9-dinydro-9-(2-thienylmetilyl)-lH ~urine-6,~3-dione lS ~ mixture of a-oro~no-9-~(2-t;~ienyl)methyL]-guanina (see Exa~ple 5? (3.12 g; 9.56 mmol), acetic anhydride (7; ml), glacial acetic acid (75 ml) an~
annydrous sodium acetate (14.9 g, 0.1816 mol) was heated to reflux for 20 hours. The ~ark solution which formed ~aJ then evaporated to dryness under reduced pressura. ~he re-idue was -lissolved in aqueous methylamine (150 ml), --tirred at room tenperature for 4a hours and then heated to reflux for 2.5 hours. The methylamine -~as ~i~tilled off under reduced prassure and the residue was racrystallize~
from ooilin~ methanol-~ater mixture to giv9 1. 23 g of the analytical pr~duct, mp ~300C.

... . . ..

- ~L28~3~3 ~DZ-l -60-~X~MPLE l~

2-~ino-l,7,8,9-tetra'nydro-9-(2-thienylmet'nyl)-~-thioxo-6~-~urin-6-one A mixture of ~-bro~o-9-C(2-thienyL)methyl]
guanine (see Exarnple 5) ~2.0 g; 6.13 mrnol), ~MF
(250 ml), and thiourea (0.93 g; 12.26 mmoL) was ~neated under r~flux for 20 hours dnd than the solvent was evaporated to dryness under reduced ~ressure. The residue was dissolved in ~J ~aOH, treated with charcoal, filtered, and acidified with ~lacial AcOH to yive a pala yellow solid. ~nalytical sample was prepared by repeatin~ the ~urifica~ion ~roces~, yield 70~ mg; mp >280C.

EX~MPL~ 20 ,~ 6,9-dih~ro-c,-oxo-9-~2-thianylmet'ny~ urin 2,8-di-yl]bis aceta.nide .~ mixture of a-~ino-9-t(2-thianyl)methyl~-guanine (0.5 g; l.88 r~ol), ~F (l0 ml), pyri~1ine (5 ~1), and ace~ic anhydri~e (5 ml) was stirred at ~room te~nperature for 36 Aours. The mi.~ture was ~ilute~ with ether (50 ml) and filtered to ~ive analytically pure product, Inp 243-~~'.

STARTI~G MAT~R~La Starting material~ are prepared a~ 'ollows usi.~g a known proce~1ure or following a ~ro~e~ure analogous o that ~own in t~e art.
5-MethyL-2-~iienyl.net.lyla~ine, H. oar-~gh, et al, J. .~n. Cnem. Soc., 19~, 70:~01~.
aenzo[b~t:'nio~en-2-yl-.methyl~nine, ~. S~.irley, 30 et ~L, J. A~. Chem. Soc., l9~2, 7~:664.

~2~3913~31 ~ EDZ-l -61-3-~ethyl-2-thienylmethylamine, H. Hartough, et al, J. ~a; Chem. Soc., 194~, 70:~01~.
BenzoLb]thio~hen-3-yl-methylamine was prepare~
by Gabriel Synthesis from the correspondin~ chloro-compound (W. King, et al, J. Org. Chem., 1948,3:635).
2-~ethyl-3-thienylmethyl~nine ~as prepared by lithium alu.-ainum hydride (LAH) reduction of the correspondin~ nitrile (M. Janda, et al, Coll. Czech.
Comm., 1974, 39:959).
2-(2-thienyl)ethyLamine and 2-(3-thienyl)ethyl-amine were preparad by the lit. method of `i. Hertz, et al, in J. ~a. Cnem. Soc., 1951, 73:351.
2-Methyl-4-thienyl methyla-aina was preparad by ~H reduction of 2-methyl-4-cyano-thiophene which was oreyarad fro,n the correspondin~ 4-oromo compound (~. Goldfarb, at al, Zh. Obs. ~nim. 1964, 34:969) and CuCN.
3-Metnyl-4-tnienylmethylaraine was s~ilarly orepared as follows:

Br ~ ~ x3 H2N-H2C ~ -C~3 ~ -Met~yL-2-~ienylaethyl~-ai.le ~as ~reoared by ~d reluction or ~he corresoondl^.g aldox~e.
2- an~ 4- T~iazolylm6r yl ~aines were ~re~ared ~ccording to the iiterat~ ~ ~roce~ur@s (~. G. Jo..es, e' al, J. ~m. Cnem. ~oc., 1950, 72:45~6).

r .~ 39~L3~

E.l~Z -1 -62-F O RMULA

~;N
<4) ~N

E~2N N~NH , III
P~r ~C~2t~ ) n ~U~NHCNHCOOR6 H2~J ~N NH II
Ar (C~2~R4 ) 5 n o J~NHCHO

2N `N~NH

.~CH2 51 n ,, . :.

Claims

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. A compound of general formula:

III) wherein:
m is zero, one, 2 or 3;
n is zero or one;
R6 represents a group selected from:
(i) phenyl, C1-4alkyl and phenyl-C1-4 alkyl, (ii) phenyl substituted by a group selected from F, Cl, Br, I, -OH, -CF3, C1-4alkyl and C1-4alkoxy, and (iii) phenyl-C1-4alkyl-ring-substituted by a group selected from F, Cl, Br, I, -OH, -CF3, C1-4alkyl and C1-4alkoxy;
R4 and R5, independently, represent a group selected from H, C3-6cycloalkyl, (i), (ii) and (iii) as defined above;
and Ar represents a group selected from:
(iv) a 5-membered aromatic ring with at least one hetero atom selected from N, O and S, (v) a 6-membered aromatic ring with at least one hetero atom selected from N, O and S, and (vi) the rings defined in (iv) and (v) substituted by a group selected from a halogen, C1-4alkyl, C1-4alkoxy and -C=C-C=C- attached to adjacent carbons to form a benzo radical;
with the proviso that at least m or n is one.
2. The compound of general formula (III) as defined in claim 1, wherein Ar represents a groups selected from 2-and 3-furanyl, 2- and 3-thienyl, 2-, 3- and 4-pyridyl, and 2-, 4- and 5-thiazolyl.
3. A process for preparing the compound of general formula (III) as defined in claim 1, comprising: reacting a compound of general formula:

(II) wherein m, n, R4, R5, R6 and Ar are as defined in claim 1, with a coupling agent, in the presence of a solvent, and recovering the desired product of general formula (III).
4. The process of claim 3, wherein the coupling agent is N,N'-dicyclohexylcarbodiimide.