USOO5272153A United States Patent (19) 11 Patent Number: 5,272,153 Mandell et al. 45 Date of Patent: Dec. 21, 1993

(54)54 OFMETH E. NHIBITINSi6. CTVTY OTHER PUBLICATIONS Avido et al., Angiology 35, 407 (1984). 75 Inventors: St. Mel ES Suguira et al., Japanese Journal of Anesthesiology 32, va.8 William,Wan, CharlotteSVlie, Novici, Lebanon, O 435-41,35-41, with English translationtranslation. N.J. (List continued on next page.) 73) Assignees: Hoechst-Roussel Pharmaceuticals, Primary Examiner-Nathan M. Nutter Inc., Somerville, N.J.; University of Attorney, Agent, or Firm-Finnegan, Henderson, Virginia Alumni Patents Foundation, Farabow, Garrett & Dunner Charlottesville, Va. (*) Notice: The portion of the term of this patent 57) ABSTRACT E. to Jun 15, 2008 has been A family of compounds effective in inhibiting interleu SC. kin-1 (IL-1) activity, tumor necrosis factor (TNF) activ (21) Appl. No.: 908,929 ity, and the activity of other leukocyte derived cyto s kines is comprised of 7-(oxoalkyl) 1,3-dialkyl xanthines (22 Filed: Jul. 2, 1992 of the formula

Related U.S. Application Data R (I) 63 Continuation of Ser. No. 700,522, May 15, 1991, aban- Y N-A-C-CH3 doned, which is a continuation of Ser. No. 622,138, Dec. 5, 1990, Pat. No. 5,096,906, which is a continua- al- 6 tion of Ser. No. 508,535, Apr. 11, 1990, abandoned, O N N which is a continuation of Ser. No. 239,761, Sep. 2, 1988, abandoned, which is a continuation of ser. No. R2 ESSEE inselected which from R1 and the R2group are consistingthe same or of different straight-chain and are or 51) int. Cl...... A61K 31/52 branched alkyl radicals with 2 to 6 carbon atoms, cyclo 52 U.S.C...... 514/263; 514/929 hexyl, alkoxyalkyl and hydroxyalkyl radicals, and A 58 Field of Search ...... 514/263,929 represents a hydrocarbon radical with up to 4 carbon w atoms which can be substituted by a methyl group. 56) References Cited Another family of effective compounds is identified as U.S. PATENT DOCUMENTS R r (II) 4,558,051 12/1985 Sunshine et al...... 514/261

4,880,791 11/1989 Weithmann et al. ... 54/26 4,965,271 10/1990 Mandell et al...... 514/929 N C) 4,975,432 12/1990 Weithmann et al...... S14/26 oes N R2 FOREIGN PATENT DOCUMENTS 0195496 9/1986 European Pat. Off. . The inhibition of IL-1, TNF, and other cytokines in 0267676 5/1988 European Pat. Off. . mammals is implicated in alleviation of a wide variety of 0344586 5/1988 European Pat. Off. . disease conditions. 0279079 8/1988 European Pat. Off. . 35 Claims, 6 Drawing Sheets 5,272,153 Page 2

OTHER PUBLICATIONS Chemical Abstracts, vol. 101, No. 19, issued Nov. 5, seret al., Journal of Leukocyte Biology, 40:747-54 1984, C.Abstract Robino No. 168805u. of Medicine, vol. 14, No. (1986). 2, 1983, Studies on Oesteoporoses.XI-Effects of Me. Chemical Abstracts, vol. 104, No. 25, issued Jun. 23, thylxanthine Drivative, pp. 137-145. 1986, Abstract No. 218828p. Clinical Aspects of White Cell Rheology Reference. U.S. Patent Dec. 21, 1993 Sheet 1 of 6 5,272,153

2.3 2. DIRECTED .9 MIGRATION (mm) 7

.5

3.

O. FIG. DBUTY OXO PROPYL XANTHENE (g/ml)

FIG 2 2.8 2.7 2.6 2.5 DIRECTED 24 MGRATION

2.2 2. 2O 9 DBOPX ug/ml U.S. Patent Dec. 21, 1993 Sheet 2 of 6 5,272,153

CONTROL FIG. 3 E. DBOPX OOlmM DBOPX mM

IOO

90

/ PMN ADHERENCE Bo

7O

6O O O. O O (DBOPX g/ml FIG. 4 U.S. Patent Dec. 21, 1993 Sheet 3 of 6 5,272,153

SUPEROXDE (nmoles per IO min million PMN)

FIG. 6

O O. O IOO U.S. Patent Dec. 21, 1993 Sheet 4 of 6 5,272,153

lugº O

$

SINNNY QNNNNNNNNNNNNNNNZZZZZZZZZZZZZ$ SSSSSSSSSSI?$ZZZZZZZZZZZZ.Ida ZZZZZZZZZZZZZZ.jszNE LPS(ZZZZZZZZ KINE U.S. Patent Dec. 21, 1993 Sheet 5 of 6 5,272,153

CELLOBJIC]NOIV89|W(uuuu) (\j-->-----C\!CN8€$$ FIG.9 KNNNNNNNNNNNNNNNNNNNC O ZZZZZZZZZZZZZZZZZZ)On [NNNNNNNNNNNN5 >>>>>>>]©ZZZZZZZZ??

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NONE LPS CONT KINES LPS KINES NCUBATION OONDITIONS U.S. Patent Dec. 21, 1993 Sheet 6 of 6 5,272,153

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7 -IO IO 3O 5O 7O 90 FIG. O L

5,272,153 1. 2

METHOD OF NHIBITING THE ACTIVITY OF O (I) LEUKOCYTE DERVED CYTOKNES R N CROSS-REFERENCE TO RELATED APPLI- 5 N N-a--ch, CATION o?als N 2 O This application is a continuation of application Ser. No. 07/700,522 filed May 15, 1991 and now abandoned, R2 which is a continuation of Ser. No. 07/622,138 filed Dec. 5, 1990 and now U.S. Pat. No. 5,096,906, which is 10 in which R1 and R2 are the same or different and are a continuation of Ser. No. 07/508,535 filed Apr. 11, independently selected from the group consisting of 1990 and now abandoned, which is a continuation of straight-chain or branched-chain alkyl radicals with 2 to Ser. No. 07/239,761 filed Sep. 2, 1988 and now aban 6 carbon atoms, cyclohexyl, straight chain or branched doned, which is a continuation of Ser. No. 06/947,905 15 chain alkoxyalkyl, and hydroxyalkyl radicals; and filed Dec. 31, 1986 and now abandoned, and is a contin A is a hydrocarbon radical with up to 4 carbon atoms uation of 07/131,785 filed Dec. 11, 1987 and now U.S. which can be substituted by a methyl group; or Pat. No. 4,965,271. a compound of the formula (II) BACKGROUND OF THE INVENTION 20 (II) This invention relates to the inhibition of activity of leukocyte derived cytokines, such as interleukin-1 and tumor necrosis factor, in humans and mammals. More specifically, this invention provides a method of inhibit ing the activity of cytokines to arrest or alleviate certain 25 disease and inflammatory states. Interleukin-1 (IL-1) and tumor necrosis factor (TNF) wherein at least one of R1 and R3 is either (a) a branched are biological substances produced by monocytes and hydroxyalkyl group of the formula other macrophages in mammals. IL-1 and TNF affect a 30 wide variety of cells and tissues, both in vitro and in vivo. Research has demonstrated that IL-1, TNF, and R4 other leukocyte derived cytokines are important, and (CH)--CH, even critical, mediators in a wide variety of inflamma OH tory states and diseases. The inhibition of IL-1, TNF, 35 and other leukocyte derived cytokines is of benefit in with a tertiary alcohol function, in which R stands for controlling, reducing, and alleviating many of these an alkyl group with 1 to 3 carbon atoms and n stands for conditions. a whole number from 2 to 5, the other R or R group Detection and inhibition of IL-1, TNF, and other that may optionally be present stands for a hydrogen leukocyte derived cytokines can be relatively easily atom or an aliphatic hydrocarbon group Rs with up to documented through in vitro analysis of polymorpho 6 carbon atoms, whose carbon chain may be interrupted nuclear neutrophil behavior. Among other activities by up to 2 oxygen atoms or may be substituted with a attributed to IL-1 and other leukocyte derived cyto hydroxy or oxo group, or (b) at least one of R or R3 is kines is the promotion of leukocyte adherence and the 45 an oxoallyl group of the formula inhibition of neutrophil chemotaxis, both directly con tributing to disease and inflammation syndromes. O Despite the desirability of inhibiting the activity of IL-1 and TNF and the activity of other leukocyte de R6-c-(CH2), rived cytokines and the ease with which inhibition can 50 wherein R6 is C1-C6 alkyl, and p=2, 3 or 4. The other be detected in vitro, there exists a need in the art for R or R3 being defined as above; and R2 represents an inhibitors of IL-1, TNF, and other cytokines, wherein alkyl group with 1 to 4 carbon atoms. The xanthine of the inhibitors are acceptable for in vivo administration. formula (I) or formula (II) is employed in an amount SUMMARY OF THE INVENTION that is effective in inhibiting the activity of IL-1, TNF, 55 and other leukocyte derived cytokines in the mammal. This invention aids in fulfilling these needs in the art Exemplary within the general formula (II), and estab by identifying a class of compounds that can be success lished as an effective IL-1 inhibitor, is the well known fully employed in alleviating conditions caused by, or and commercially available pharmaceutical pentoxifyl mediated by, IL-1, TNF, and other leukocyte derived 60 line. Although this compound has been used for some cytokines. The compounds exhibit marked inhibition of time as a pharmaceutical (clinical trials in 1971) it has cytokine activity, even at low concentrations of the not been reported effective as an IL-1 inhibitor. It has mediators as demonstrated through in vitro tests. been demonstrated in promoting directed migration of More particularly, this invention provides a method leukocytes. of inhibiting the activity of IL-1, TNF, and other leuko- 65 cyte derived cytokines in a mammal comprising admin BRIEF DESCRIPTION OF THE DRAWINGS istering thereto at least one 7-(oxoalkyl) 1,3-dialkyl This invention will be more fully described with xanthine of the formula (I) reference to the drawings in which: 5,272,153 3 4. FIG. 1 is a graph showing modulation by 1,3-dibutyl zyme secretion by macrophages, and factors that medi 7-(2-oxopropyl) xanthine (DBOPX) of the effect of ate secretion of oxidizing agents, such as oxygen, super interleukin-1 (IL-1) on polymorphonuclear leukocyte oxide, hydrogen peroxide, and hydroxyl radical. (PMN) directed migration to n-formylmethionyl leucyl Xanthines employed in this invention have the fol phenylalanine (FMLP); 5 lowing formula: FIG. 2 shows the results of modulation by DBPOX of the effect of mononuclear leukocytes LPS stimulated conditioned medium on PMN directed migration to FMLP; O N N-A-C-CH3 FIG. 3 shows the results of modulation by DBOPX H of the effect of tumor necrosis factor (TNF) on PMN oals N 2 O directed migration to FMLP; N FIG. 4 shows the results of modulation by DBOPX R2 of LPS stimulated mononuclear leukocyte conditioned 15 medium on PMN adherence to nylon; The substituents R1 and R2 in formula (I) are the same or FIG. 5 shows the results of modulation by DBOPX different and are independently selected from the group of IL-1 on PMN superoxide release stimulated by consisting of straight-chain or branched alkyl radicals FMLP; with 2 to 6 carbon atoms, cyclohexyl, alkoxyalkyl and FIG. 6 is a graph showing modulation by DBOPX of 20 lipopolysaccharide (LPS) stimulated mononuclear leu hydroxyalkyl radicals. The substituent A represents a kocyte conditioned medium on superoxide production hydrocarbon radical with up to 4 carbon atoms, which by PMN stimulated with FMLP; can be substituted by a methyl group. FIG. 7 is a graph showing modulation by DBOPX of A compound that has been found to be particularly the effect of LPS-stimulated mononuclear leukocyte effective for inhibiting the effects of IL-1 and other conditioned medium on lysozyme released by PMN leukocyte derived cytokines on polymorphonuclear stimulated with FMLP; leukocytes and monocytes is 1,3-dibutyl 7-(2-oxopro FIG. 8 shows modulation by pentoxifylline of the pyl) xanthine. This compound, which is also referred to effect of LPS stimulated mononuclear leukocyte condi- 30 herein in abbreviated form as "DBOPX', has the fol tioned medium on PMN directed migration; lowing formula: FIG. 9 shows modulation by pentoxifylline of the effect of C. albicans stimulated mononuclear leukocyte conditioned medium on PMN directed migration; O (III) FIG. 10 shows modulation by pentoxifylline of the 35 effect of IL-1 on PMN directed migration; CHm(CH3-( R): N FIG. 11 shows modulation by pentoxifylline of the effect of LPS stimulated mononuclear leukocyte condi 2 N tioned medium on PMN adherence; and o? N FIG. 12 shows modulation by pentoxifylline of the (CH2)3CH3 effect of IL-1 and LPS stimulated mononuclear leuko cyte conditioned medium on PMN adherence. “DBOPX' DESCRIPTION OF THE PREFERRED The ability of compound (III) to inhibit the effects of EMBODIMENTS 45 IL-1 and other leukocyte derived cytokines on poly Inhibition of the activity of IL-1, TNF, and other morphonuclear leukocyte and monocyte adherence, leukocyte derived cytokines can be achieved by the cell chemotaxis, respiratory (metabolic) burst, and cell administration of xanthines of formula (I) or formula degranulation has been demonstrated and is described (II) to a mammal. 50 hereinafter. As used herein, the expression "leukocyte derived Inhibition of IL-1 activity can also be achieved by the cytokines" is to be given a broad meaning. Specifically, administration of compounds of the formula (II) the term "leukocyte' as used herein means mammalian cells of granulocytic and lymphocytic lineage. Exam ples of leukocyte cells are polymorphonuclear leuko- 55 R O R3 cytes, such as neutrophils, and mononuclear phago cytes, such as monocytes and macrophages, and lym A. phocytes. oals) N N The term "cytokine' as used herein means a secre tory product of a leukocyte, and in particular a non k antibody protein released by a leukocyte on contact with antigen and which acts as an intercellular mediator to the host or patient to be treated. As noted, among of immune response. Examples of cytokines that are these compounds is the commercially available pentoxi within the scope of this invention are chemotactic fac- 65 fylline. A host of other compounds within the general tors, factors promoting replication of lymphocytes, formula (II) have been identified as demonstrating IL-1 factors inhibiting replication of lymphocytes, factors inhibiting activity. Among these compounds are those affecting macrophage adherence, factors affecting en identified by their R substituents set forth below. 5,272,153 5

COMPOUND OF FORMULA (II) Compound if R R2 R3 2 -CH3 -CH2-CH2-CH3 CH3-C-(CH2)4- 3 ph -CH3 -CH2-CH2-O-CH3 CH--(CH CH3

4. PP -CH2-O-(CH2)2-O-CH 5 p AF -H 6 P -CH2-CH2-CH3 7 p ph -CH2-CH-CH3 8 PF gh -CH2-CH-(CH3)2 9 -CH2-CH3 -CH2-O-CH-CH 10 -CH3 CH3 -(CH2--CH, OH -CH2-O-CH2-CH3 When introduced into polymorphonuclear neutrophil The migration of polymorphonuclear leukocytes in (PMN) incubations provided with IL-1, or incubated in response to n-formyl methionyl leucyl phenylalanine lipopolysaccharide stimulated mononuclear leukocyte (FMLP), a well known chemotactic factor, was deter condition medium, the compounds of formula (II) of the mined by chemotaxis under agarose, a well known assay claimed invention decreased PMN adherence, even at 35 for cell chemotaxis. See J. of Immunol, 115, 6, relatively low concentrations (0.1 of micrograms/ml). 1650-1656 (1975). The assay was carried out without Similarly, the presence of the compounds of formula IL-1, and the assay was repeated in the presence of (II) of the claimed invention promoted directed migra IL-1. The assay was also carried out with IL-1, but tion of PMN, which migration is inhibited by the pres without DBOPX, and with both IL-1 and DBOPX at ence of IL-1. The demonstrated inhibition of IL-1 by DBOPX concentrations of 0.1, l, and 10 micrograms these compounds is, of course, suggestive of clinical per milliliter (ug/ml). The results are depicted in FIG. effectiveness in the above-identified areas, and addi 1. tional conditions. Appropriate dosages will vary with As shown in FIG. 1, directed migration of the cells in the condition and individual. the absence of IL-1, TNF, and with 0 g/ml DBOPX Phagocytes important in immunology are polymor 45 (i.e. "CONT" in FIG. 1) was about 2.08 mm. Directed phonuclear leukocytes (e.g. neutrophils) and migration of the cells dropped to about 1.5 mm in the mononuclear phagocytes (e.g. monocytes and macro presence of IL-1, TNF, and with 0 g/ml DBOPX. phages). Phagocyte hypofunction is a cause of recurrent Thus, IL-1 inhibited cell chemotaxis directed to FMLP. pyogenic infection. To combat pyogenic infection, neu FIG. 1 also shows the effect of increasing concentra trophils and monocytes respond to chemotactic factors 50 tions of DBOPX on the inhibition of chemotaxis by by moving toward the source of infection, where they IL-1. More particularly, DBOPX modulates the inhibi ingest microorganisms and kill them. tory effect of IL-1 on directed migration to FMLP. More particularly, a main function of polymorphonu Specifically, FIG. 1 shows that DBOPX increased di clear leukocytes and monocytes is to kill bacteria and rected migration of the cells and modulated the inhibi other infectious agents by phagocytosis. The first stage 55 tory effect of IL-1 at all of the DBOPX concentrations in the ingestion and digestion of a particulate substance that were evaluated. FIG. 1 also shows that DBOPX by these cells involves the process of bringing the cells was effective in increasing chemotaxis even at very low and the particles together, usually through chemotaxis. DBOPX concentrations. Thus, the compounds em This response is an essential part of host defence against ployed in the process of this invention are particularly infection. The extensive migration and activity of these effective in modulating the inhibitory effect of IL-1 on cells is manifested by inflammation at the site of injury cell chemotaxis. or invasion of the host. DBOPX is capable of producing a similar effect on it has been shown that IL-1 and TNF inhibit chemo polymorphonuclear leukocytes incubated with the taxis by granulocytes, monocytes and macrophages. It products of mononuclear leukocytes that were stimu has now been discovered that the 7-(oxoalkyl) 1,3-dial 65 lated with lipopolysaccharide (LPS). These kyl xanthines of formula (I) are capable of modulating mononuclear cells produce IL-1, TNF, and other in the inhibitory effect of IL-1 and TNF on chemotaxis. flammatory cytokines. Once again, polymorphonuclear This has been demonstrated as follows. leukocyte directed migration to FMLP was determined 5,272,153 7 8 by chemotaxis under agarose. The assay was carried out herence. At a DBOPX concentration of 10 (ug/ml) the without DBOPX and with concentrations of DBOPX percent PMN adherence declined to about 70%. The of 0.1, 1.0, 10, and 100 g/ml. The results are shown in probability that DBOPX decreased adherence of PMN FG, 2. incubated with conditioned medium was 99.7%. Thus, Referring to FIG. 2, the directed migration of the 5 the compounds employed in the process of this inven PMN in the conditioned medium containing the inflam tion are particularly effective in blocking adherence of natory cultures was about 2.25 mm in the absence of leukocytes and thereby aiding in reducing the degree of DBOPX. The addition of DBOPX to the medium in inflammation. creased directed migration of the cells at all of the Mature phagocytes are in a metabolically dormant DBOPX concentrations tested. Once again, DBOPX 10 state. It is currently believed that recognition of certain was effective in increasing chemotaxis even at very low objects and substances by phagocytes, such as the at concentrations. Moreover, the directed migration was tachment of an ingestible particle to the cell surface, about 2.6 mm at a DBOPX concentration of 10 g/ml. changes this situation, and the cell enters a stage of By comparison, migration in an unconditioned medium increased metabolic activity, which is referred to as containing LPS was 2.600.5 mm. (Data not shown in 15 metabolic or respiratory burst. The transition is associ FIG. 2). The probability that DBOPX increased di ated with a series of characteristic changes, including rected migration inhibited by conditioned medium con the production of a superoxide anion. Cytokines, such taining inflammatory cultures was 95%. as IL-1 and TNF, are capable of producing a similar DBOPX is capable of producing a similar effect on effect. In addition to its significance for phagocytic PMN incubated with rhi-TNF (alpha). PMN directed 20 function related to inactivation of ingested microbes, migration to FMLP was determined by chemotaxis activation of oxygen metabolism is a useful indirect under agarose. The assay was carried out without marker for the ingestion process per se. It would be DBOPX and with concentrations of DBOPX of 0.01 desirable to be able to modulate the effect of cytokines mM (3.2 g/ml) and 1 mM (320 g/ml). The results are on respiratory burst. shown in FIG. 3. 25 Quantitative methods for direct measurement of hy Referring to FIG. 3, the directed migration of the drogen peroxide and superoxide anions released into the PMN in medium containing rh-TNF was 1.45mm in the medium are currently available. It has been found that absence of DBOPX. The addition of DBOPX to the the compounds employed in this invention are capable medium increased directed migration of the cells at both of modulating respiratory burst in stimulated polymor of the DBOPX concentrations tested. Once again, 30 phonuclear leukocytes (PMN) as determined using DBOPX was effective in increasing chemotaxis even at these methods. very low concentrations. By comparison, migration in More particularly, superoxide production was as medium in the absence of TNF was 2.75 mm. The prob sayed using a modification of the procedure described ability that DBOPX increased directed migration inhib by Babior et al., J. Clin. Investigation, 52:741-744 (1973). ited by TNF was better than 95%. 35 Purified PMN were incubated with an oxidative stimu Thus, the 7-(oxoalkyl) 1,3-dialkyl xanthines em lus with and without IL-1. The medium was assayed for ployed in the process of invention are capable of in superoxide production. The assay was also carried out creasing directional movement of polymorphonuclear without DBOPX and with DBOPX in concentrations leukocytes. These compounds can be administered to a of 0.1, 1.0, 10, and 100 g/ml. The results are shown in patient to augment chemotactic factors of bacterial or 40 FIG.S. viral origin, or components of plasma activation sys It is evident from FIG. 5 that about 1.8 nmoles of tems, or factors elaborated by cells of the immune sys superoxide/10 min/million PMN were produced by ten, FMLP-stimulated PMN in the absence of IL-1, TNF, Leukocyte response to an acute inflammatory stimu and DBOPX (see "CONT" in FIG. 5). Pretreatment lus involves a complex series of events, including adher 45 with IL-1 (5 units/20 Jul), which is known as priming, ence to endothelium near the stimulus. Inhibition of produced a substantial increase in observed superoxide leukocyte adherence can be expected to reduce the release to about 4.4 nmoles superoxide/10 min/million degree of inflammation seen in conditions, such a septic PMN. shock and adult respiratory distress syndrome. It has In contrast, the addition of DBOPX to the assay been found that the 7-(oxoalkyl) 1,3-dialkyl xanthines SO resulted in a substantial reduction in observed superox employed in this invention effectively block adherence ide production as is evident from FIG. 5. Specifically, of polymorphonuclear leukocytes. DBOPX modulated the effect of IL-1 on stimulated Specifically, polymorphonuclear leukocyte (PMN) PMN at all of the concentrations tested. DBOPX was adherence to nylon was determined according to the even effective at a very low concentration of 0.1 g/ml. method of MacGregor et al., New Engl. J. Med. 55 The probability that DBOPX decreased superoxide 13:642-646 (1974). Purified PMN cells were incubated production produced by PMN primed with IL-1, TNF, with a lipopolysaccharide-stimulated mononuclear leu and stimulated with FMLP compared with IL-1 alone kocyte conditioned medium containing inflammatory was 95%. cytokines. PMN adherence to nylon was determined DBOPX is also capable of decreasing superpxide without DBOPX, and then with DBOPX at concentra production by PMN primed with LPS-stimulated tions of 0.1, 1.0, and 10 g/ml. The percent PMN ad mononuclear leukocyte conditioned medium containing herence to nylon was determined for each case. The inflammatory cytokines. This is shown in FIG. 6. Spe results are summarized in FIG. 4. cifically, when PMN were incubated with LPS FIG. 4 shows that PMN adherence to nylon in the stimulated mononuclear leukocyte conditioned medium absence of DBOPX was about 87%. However, when 65 containing inflammatory cytokines and stimulated with DBOPX was included in the assay at concentrations FMLP, observed superoxide production in the absence above about 0.1 (ug/ml) PMN adherence to the nylon of DBOPX was about 7.4 nmoles/10 min/million PMN. was inhibited as evidenced by a decline in percent ad When DBOPX was added to the assay, however, ob 5,272,153 10 served superoxide production was lower at all of the cells. The compounds can decrease oxidative damage to DBOPX concentrations tested. Moreover, DBOPX host tissues by phagocytes as evidenced by modulation exhibited some effect even at a concentration as low 1.0 of respiratory burst in stimulated polymorphonuclear ug/ml. At a DBOPX concentration of 10 g/ml, super leukocytes. Finally, the compounds can modulate the oxide production was about 1.5 nmoles/10 min/million effects of cytokines on degranulation in stimulated PMN. The probability that DBOPX decreased superox phagocytes. The demonstrated inhibition of IL-1, TNF, ide production produced by PMN primed with condi and other cytokines by these compounds is suggestive tioned medium and stimulated with FMLP was 99.5%. of clinical effectiveness in at least the following areas It is evident from these results that the compounds and conditions. employed in the process of this invention are capable of 10 reducing superoxide production and modulating respi Because IL-1, TNF, and other leukocyte derived ratory burst in phagocytes, such as polymorphonuclear cytokines have been implicated in such a wide variety leukocytes and monocytes. of mammalian conditions, this invention has a similarly During ingestion, granules in the cytoplasm of the broad scope of application. Among the conditions that cell fuse with the membrane of a vacuole that was 15 can be treated or alleviated by the inhibition of IL-1, formed around the foreign substance. The granules TNF, and other leukocyte derived cytokines are: sepsis, discharge their contents into the vacuole. Some of this septic shock, endotoxic shock, gram negative sepsis, material ends up in the medium surrounding the phago toxic shock syndrome, adult respiratory distress, fever cyte. Since the granules disappear during this process, it and myalgias due to infection (i.e. influenza), cachexia is called degranulation. The granule contents include 20 secondary to infection or malignancy, cachexia second hydrolytic enzymes, lysozyme, bactericidal proteins, ary to AIDS, rheumatoid arthritis, gouty arthritis, oste and, in the neutrophil, myleoperoxidase. oporosis, keloid formation, scar tissue formation, de Degranulation can be assessed by measuring the rate creased appetite, Crohn's disease, ulcerative colitis, of appearance of granule-associated enzymes in the fever due to central nervous system bleeding, glomeru extracellular medium. In the case of polymorphonu 25 lonephritis, multiple sclerosis, Creutzfeld-Jacob disease, clear leukocytes (PMN), degranulation can be assayed adverse reactions to dialysis, diabetes melitus, and psori by determining release of lysozyme. It was found that S.S. the compounds employed in the process of this inven By reference to the specific cause of the disease con tion are capable of modulating the release of lysozyme dition, the more generic term "trauma' can be used. from stimulated PMN. 30 The term "trauma' refers broadly to cellular attack by More particularly, polymorphonuclear leukocytes foreign bodies and physical injury of cells. Included (PMN) were incubated with LPS-stimulated among foreign bodies are microorganisms, particulate mononuclear leukocyte conditioned medium containing matter, chemical agents, and the like. Included among inflammatory cytokines. The PMN were then stimu physical injuries are mechanical injuries, such as abra lated with FMLP, incubated for a period of time, and 35 sions, lacerations, contusions, wounds, and the like; lysozyme content was determined in cell supernatant therma injuries, such as those resulting from excessive using a well known assay. See J. Bacteriol, 58,731-736 heat or cold; electrical injuries, such as those caused by (1949). The PMN were incubated without DBOPX or contact with source of electrical potential; and radiation with DBOPX in a concentration of 0.1, 1, 10, or 100 damage caused, for example, by prolonged, extensive ug/ml. The results, which are expressed in terms of 40 exposure to infrared, ultraviolet or ionizing radiations. lysozyme released/10 min/4 million PMN (ug/ml), are Microorganisms included among the foreign bodies shown in FIG. 7. that can elicit a biological response are bacilli, fungi and Referring to FIG. 7, lysozyme released by PMN yeast, viruses, parasites, and the like. Representative primed with LPS-stimulated mononuclear leukocyte bacilli are: Actinomyces spp.; Bacteroides spp.; Coryne conditioned medium (containing inflammatory cyto 45 bacterium spp.; Enterobacteriacea; Enterococcus; Hae kines) and stimulated with FMLP was about 2.1 g/ml mophilus spp.; Micrococcus spp.; Neissera spp.; Staphy in the absence of DBOPX. When DBOPX was added to lococcus aureus, Streptococcus pneumoniae; Clostridium the assay, lysozyme release declined. The decrease was spp.; Streptococcus agalactiae; Bacillus spp.; H. influen observed at all of the concentrations of DBOPX that zae; Moraxella spp.; Mycobacteria spp.; Pseudodomonas were evaluated. Moreover, DBOPX was effective in 50 aeruginosa; Vibrio spp.; and Mycoplasma. modulating lysozyme release even at concentrations as Representative fungi and yeast that are capable of low as 0.1 g/ml. At a DBOPX concentration of 100 eliciting a biological response are: Microspurum; Blas ug/ml, the lysozyme release was only about 1.04 tomcyes; Histoplasma; Aspergillus; Cryptococcus; Can ug/ml. The probability that DBOPX inhibited lyso dida spp.; Coccidioides; and Candida albicans. zyme release from PMN primed with conditioned me 55 Representative viruses are: Rhinovirus; Parainflu dium and stimulated with FMLP was 95%. enza; Enterovirus; Influenza; Smallpox and vaccinia; It is apparent from these results that the compounds Herpes simplex; Measles; Rubella; Arbovirus (Western, employed in the process of this invention are capable of Eastern and Venezuelan equine encephalitis, and Cali decreasing the release of lysozyme from PMN primed fornia encephalitis); Rabies; Colorado tick fever; Yel with LPS-stimulated mononuclear leukocyte condi 60 low fever; Dengue; Hepatitis Virus B (HB Ag); Hepati tioned medium and then stimulated with FMLP. tis Virus A (HAV); and Human Immunodeficiency In summary, the compounds of formula (I) employed Virus (HIV). in the process of this invention are capable of modulat Representative parasites that can elicit a response are: ing the effects of leukocyte derived cytokines, such as Trypanosoma cruzi Entamoeba histolytica, Leishmania interleukin-1 and tumor necrosis factor, on phagocytes, 65 brasiliensis, Leishmania tropica, Leishmania donovani such as polymorphonuclear leukocytes. The com Toxiplasma gondii Plasmodium falcipaum, Trypanosoma pounds are capable of substantially aiding chemotaxis. rhodesiense Loa loa, Trichomonas hominis, Schistosoma In addition, the compounds can block adherence of japonicum, Schistosoma mansoni; and Fasciola hepatica, 5,272,153 11 12 Particulate materials capable of eliciting a biological defined above, with oxoalkyl halides corresponding to response include silica, asbestos, monosodium urate, the formula cotton fibers coal dust, beryllium, and the like. Chemical agents include heavy metals, such as lead, CH3-C-A-Hal (VI) chromium, mercury, arsenic, and the like; organic sol I vents, such as trichloroethylene, and the like; herbi O cides, such as trichlorophenoxyacetic acid and the like; and pesticides, such as mirex and the like. in which A is as defined above, and Hal represents a In addition, inhibition of IL-1, TNF, and other leuko halogen atom, preferably chlorine or bromine. cyte derived cytokines will enhance phagocyte activity 10 These reactions are preferably carried out attempera in stored blood and blood products. tures in the range from 40" to 80' C., optionally under The compounds employed in this invention will now elevated or reduced pressure, but usually at atmo be described in more detail, and methods for preparing spheric pressure. The individual starting compounds the compounds will be provided. can be employed either in stoichiometric quantities or in The process of this invention utilizes 7-(oxoalkyl) 15 excess. The alkali salts in the alternative method of 1,3-dialkyl xanthines of formula (I) above. While preparation can either be prepared beforehand or in the DBOPX is the particularly preferred xanthine, a num reaction itself. , ber of other compounds can be employed. For example, Suitable solvents for use in the reactions are water the xanthines of formula (I) can be substituted by other miscible compounds, preferably lower alcohols, such as alkyl groups, or by alkoxy or hydroxyalkyl groups. 20 methanol, propanol, isopropanol, and various butanols; Suitable alkyl groups include branched and straight also acetone; pyridine; triethylamine; polyhydric alco chain groups, such as ethyl, propyl, isopropyl, butyl, hols, such as ethylene glycol and ethylene glycol mono sec-butyl, tert-butyl, amyl, hexyl, and the like. Alkoxy methyl or monoethyl ether. substituted alkyl groups are branched and straight chain The compounds of formula (I) are known for their groups containing from 2 to 6 carbon atoms in the com 25 marked effect in increasing blood flow through skeletal bined alkoxy and alkyl groups, including methox muscle and by their low toxicity. The most active of ymethyl, amyloxymethyl, methoxyethyl, butoxyethyl, these compounds for use in accordance with the present propoxypropyl, and the like. Hydroxyalkyl groups are invention is 1,3-dibutyl 7-(2-oxopropyl)xanthine, i.e. those containing from 1 to 6 carbon atoms, such as DBPOX. hydroxymethyl, hydroxyethyl, hydroxypropyl, hy 30 A more detailed description of the compounds of droxyhexyl, and the like. formula (I) employed in this invention and methods of The hydrocarbon groups represented by A in for preparing the compounds are contained in U.S. Pat. No. mula (I) above are divalent saturated aliphatic hydro 4,242,345, the entire disclosure of which is relied upon carbon groups, i.e., methylene, ethylene, trimethylene and incorporated by reference herein. and tetramethylene, which can be substituted on the 35 As noted, among the compounds of formula (II) en carbon adjacent the carbonyl group with methyl. Such braced in this invention is pentoxifylline (Trental (R)). methyl-substituted groups include ethylidine, 1,2-pro Other compounds can be prepared according to the pylene, and 1,3-butylene groups. disclosure of U.S. Pat. No. 3,737,433 and Belgium Pa The compounds of formula (I) employed in this in tent 831,051 (where R/R3 are oxoallyl). For the cases vention can be synthesized using known techniques. where at least one of R/R3 is a tertiary alcohol refer For example, the compounds can be prepared at ele ence may be had to the international application PCT vated temperature, optionally in the presence of a sol EP-86-00401, Jul. 8, 1986 claiming German priority of vent, by reacting correspondingly substituted 1,3-dial Jul. 8, 1985. This application addresses, as its invention, kyl xanthines of the formula 45 a variety of embodiments of synthesis routes for the xanthines of formula (II) embraced in the current inven O H (IV) tion. *l J An example of one embodiment consists of N a) reacting 3-alkylxanthines of formula (VII) 50 oals N N w (VII) , HQ \" in which R1 and R2 are as defined above, with af3 55 ossOC) N unsaturated methyl ketones corresponding to the for mula R2 in which the R3 represents alkyl with up to 4 carbon HCC-C-CH3. (V) atoms, with alkylating agents of formula (VIII) O R4 (VIII) The substituent Rin formula (V) represents hydrogen or a methyl group. The reaction can be conducted in an alkaline medium. 65 OH An alternative method of preparation involves react ing alkali metal salts of 1,3-dialkyl xanthine derivatives in which X stands for halogen, preferably chlorine, of general formula (IV), in which R1 and R2 are as bromine, or iodine, or a sulfonic acid ester group or a 5,272,153 13 14 phosphoric acid ester group and R and n have the meanings mentioned above, to obtain compounds of Rl5-X (XIII) formula (IX) with the formation of 3,7-disubstituted xanthines of R3 (IX) 5 formula (XIV) O (CH --ch HQ / 2) 3 O R6 (XIV) N N OH HQ / N N OSC) N N in O N C)N k k with a tertiary hydroxyalkyl group in the position of R3 15 in which R1 has the meaning mentioned for R or and hydrogen in the position of R, and stands for benzyl or diphenylmethyl, and then substitut al) alkylating this with the same or different alkylat ing them in the 1-position, again preferably in the pres ing agent of formula (VIII) to obtain compounds pursu ence of basic media or in the form of their salts, with a ant to the invention of formula (X) compound of formula (VIII), with compounds of for 20 mula (XV) ' r" (X) R4 (XV) Hic--(CH.) CH)--CH O R15 OH N C)A. OH Hic--(CH) oes NN 25 OH N C)M R2 oes an R2 with two identical or different tertiary hydroxyalkyl 30 groups in the positions of R and R, or being obtained, and converting the compounds of for a2) converting it with a compound of the formula mula (XV) in which R15 represents a benzyl or diphe nylmethyl being obtained, and converting the com R-X OXa) pounds of formula (XV) in which R5 represents a ben 35 zyl or diphenyimethyl group or an alkoxymethyl or in which X has the meaning given in formula (VIII) and alkoxyalkoxymethyl group, under reducing or hydro Rhas the meaning indicated above, into compounds of lytic conditions, into compounds pursuant to the inven formula (XI) tion of formula (XVI)

R4 (XI) 40 R4 (XVI) O R5 O / (CH2)2)n –c4ch 3 H3C-C-(CH2) I h N N Y N OH OH N ) 45 ) o-N N N o? N N k k in all cases preferably operating in the presence of basic that are subsequently reacted again, if desired, with a media or using the xanthines in the form of their salts. 50 compound of formula (VIII) or (Xa) to obtain com Another form of embodiment b) consists of substitut pounds pursuant to the invention of formula (X) or ing 1,3-dialkylated xanthines of formula (XII) (XV). Another form of embodiment d) consists of reducing compounds of formula CXI) or (XV) pursuant to the r: R (XII) 55 invention in which R5 or R15 stands for an oxoalkyl group, with conventional reducing agents for the keto A. group to obtain the corresponding hydroxyalkylated o N C)N xanthines pursuant to the invention. The 3-alkyl- or 1,3-dialkylxanthines of formula (VII) 60 or (XII) used here as starting materials and the "alkylat in the a-position, preferably in the presence of basic ing agents' of formulas (VIII), (Xa), and (XIII) are media or in the form of their salts, by one-step reaction known for the most part or can be prepared readily by with a compound of formula (VIII), to obtain com methods disclosed in the literature. Thus, the tertiary pounds of formula (XI). alcohols of formula (VIII), for example, can be obtained Another form of embodiment c) consists of first re 65 by organometallic synthesis by reacting the sterically acting the 3-alkylxanthines of formula (VII), likewise preferably in the presence of basic media or in the form unhindered haloketones of the formula of their salts, with a compound of the formula Hall-(CH2)n-CO-CH3 (XVII) 5,272,153 15 16 (a)-1)-hydroxy-(a)-1)-methylbutyl, -pentyl, -hexyl, and in a so-called synthetic reaction with reductive alkyla -heptyl, the (a)-2)-hydroxy-(o-2)-methylpentyl, tion of the carbonyl group, with alkylmetal compounds -hexyl, -heptyl, and -octyl, and the (a)-3)-hydroxy-(a)- R-M, especially of magnesium, zinc, or lithium, for 3)-methylhexyl, -heptyl, -octyl, and -nonyl chlorides, example in the form of alkylmagnesium halides R bromides, iodides, sulfonates, and phosphates. MgHal (Grignard compounds) or of the alkyllithium Among the compounds of formula R-X (Xa) or compounds R-Li under the usual conditions (for exam R15-X (XIII) suitable for the introduction of R into ple, see Houben-Weyl, Vol. VI/1 a, Part 2 (1980), pp. the 1- or 7-position and of R1 into the 7-position of the 928-40, especially pp. 1021 ff. and 1104-1112). In the xanthine skeleton, the alkoxymethyi and alkoxyalkox same way, a reaction of the haloketones with the for 10 mula ymethyl derivatives occupy a special position as their halides can indeed be used successfully as reactants but Hal-(CH2)CO-R' (XVIII) toxicological problems can arise, at least in large-scale use. For this reason, the use of the corresponding sulfo with methylmagnesium halides or methyllithium like nates is preferred in this special case, which are readily wise leads to the target. 15 available, for example, by reacting mixed anhydrides of The hydroxyketones corresponding to the formulas aliphatic carboxylic acids and aliphatic or aromatic (XVII) and (XVIII) can also be converted smoothly sulfonic acids (M. H. Karger et al., J. Org. Chen. 36 into diols with the alkylmetal compounds in the usual (1971), pp. 528-531) with the formaldehyde dialkyl way, either directly or with temporary masking of the acetals or dialkoxyalkyl acetals in a smooth and nearly hydroxy group, for example by acetal formation with 20 quantitative reaction (M. H. Karger et al., J. Amer. 5,6-dihydro-4H-pyran (for example, see Houben-Weyl, Chem. Soc. 91 (1969), pp. 5663/5665: Vol. VI/l a, Part 2 (1980), pp. 1113-1124), from which compounds of formula (VIII) are formed by selective R7-SO-O-CO-(C1-C4)Alkyl + esterification of the terminal primary hydroxyl groups with sulfonyl or phosphoric halides or anhydrides, ad 25 vantageously in the presence of basic media. R-o-CH-O-R-GR7-so-o-CH-O-R Other possibilities for the synthesis of the tertiary alcohol derivatives of formula (VIII) consist of the In this equation, R7 represents an aliphatic group such monometallation of co-chloro-1-bromoalkanes to obtain as methyl, ethyl, or trifluoromethyl, or an aromatic a-chloroalkylmetal compounds, (Houben-Weyl, Vol. 30 group, for example, phenyl, 4-tolyl, or 4-bromophenyl, XIII/2 a (1973), pp. 102 and 319) and their subsequent but preferably methyl or 4-tolyl, and R represents an reaction with the ketones R-CO-CH3, with the alkyl or alkoxyalkyl group falling under the definition extent of by-product formation from the alkanolates of R5 or R15. formed as intermediates because of their tendency The reaction can be carried out either in the sub toward ring closure with the elimination of metal salt 35 stance or in an anhydrous aprotic solvent inert to the being minimized by appropriate temperature control, or reactants at temperatures between -20' and +40' C., of using co-halo-l-alkanols as starting materials, which preferably between 0' and 20' C. No intermediate isola are metallated in the usual way, preferably in the form tion of the highly reactive sulfonates, which are sensi of the tetrahydropyranyl-(2) ether or after alkanolate tive to hydrolysis and thermally labile, is necessary; formation of the hydroxy group (MO-(CH2)n-Hal) 40 they are preferably used immediately as crude products with any desired alkylmetal compound (for example, for the substitution on the nitrogen of the xanthines, see Houben-Weyl, Vol. XIII/2 a (1973, p. 113), then with the usual addition of a basic condensing agent reacting them with the ketones R-CO-CH3 to obtain being unnecessary. the diols mentioned in the preceding paragraph (Houb The reaction of the mono- or disubstituted xanthine en-Weyl, Vol. VI/1 a, Part 2 (1980), p. 1029), and subse 45 quently selectively esterifying the primary hydroxy derivatives, (IX), (XVI), (VII), (XII), and (XIV) with group with suitable sulfonic or phosphoric acid deriva the alkylating agent involved of formula (VIII) or (Xa) tives. or (XIII) is ordinarily done in a distributing agent or A convenient access to compounds of formula (VIII) solvent inert to the reactants. Practical representatives in which R' represents a methyl group is also available SO are especially dipolar, aprotic solvents, for example through the reaction of a-haloalkanoic acid alkyl esters formamide, dimethylformamide, dimethylacetamide, (Hal-(CH2)n-COO-alkyl) with two equivalents of a N-methylpyrrolidone, tetramethylurea, hexamethyl methylmetal compound, with the ester reacting phosphoric triamide, dimethyl sulfoxide, acetone, or through the ketone to produce the tertiary alcohol with butanone; however, alcohols such as methanol, ethyl the introduction of two methyl groups (Houben-Weyl, 55 ene glycol, and their mono- or dialkyl ethers with the Vol. VI/1 a, Part 2 (1980), pp. 1171-1174). In the same alkyl group having 1 to 4 carbon atoms but both to way, c)-hydroxycarboxylic acid esters can be converted gether having a maximum of 5 carbon atoms, ethanol, into diols with methylmetal compounds with or without propanol, isopropanol, and the various butanols; hydro protection of the hydroxy group, for example in the carbons such as benzene, toluene, or xylenes; haloge form of tetrahydropyranyl-(2) or methoxymethyl ether, 60 nated hydrocarbons such as dichloronethane or chloro or optionally in the form of the lactones as cyclic esters form; pyridine, and mixtures of the solvents mentioned (for example, see Houben-Weyl, Vol. VI/1 a, part 2 or their mixtures with water can also be used. (1980), pp. 1174-1179), from which active alkylating The "alkylation reactions' are suitably carried out in agents of formula (VIII) can in turn be obtained by the presence of a basic condensing agent. Examples of selective esterification of the primary hydroxyl group 65 materials suitable for this are alkali metal or alkaline with sulfonic or phosphoric halides or anhydrides, earth hydroxides, carbonates, hydrides, alcoholates, and Suitable compounds of formula (VIII) that can be organic bases, such as trialkylamines (for example, prepared by the methods described above are thus the triethyl- or tributylamine), quaternary ammonium or 5,272,153 17 18 phosphonium hydroxides and crosslinked resins with fixed, optionally substituted ammonium or phospho The 1,3,7-trisubstituted xanthines of formula OXV) nium groups. The xanthine derivatives can also be used that have an alkoxymethyl or alkoxyalkoxymethyl in the alkylation reaction directly in the form of their group in the position of R represent O,N-acetals. Con separately prepared salts, such as the alkali metal, alka sequently, their substituents in the 7-position can be split line earth, or optionally substituted ammonium or phos off under the usual conditions of acid hydrolysis (cf. phonium salts. The mono- and disubstituted xanthine Houben-Weyl, Vol. VI/I b (1984), pp. 741-745), with derivatives can also be alkylated either in the presence of the aforementioned inorganic condensing agents or the 7H compounds of formula (XVI) likewise being in the form of their alkali metal or alkaline earth salts 10 formed. Examples of preferred groups that can be elimi with the assistance of so-called phase transfer catalysts, nated hydrolytically are the methoxy, ethoxy, and pro for example tertiary amines, quaternary ammonium or poxymethyl groups as well as the methoxyethoxy- and phosphonium salts. or crown ethers, preferably in a ethoxyethoxymethyl groups. The reaction is advanta 2-phase system under the conditions of phase transfer geously carried out with heating in dilute mineral acids catalysis. Among the suitable phase transfer catalysts 15 such as hydrochloric or sulfuric acid, optionally with that are generally commercially available are tetra(C- 1-C4)alkyl- and methyltrimethylammonium and -phos the addition of glacial acetic acid, dioxane, tetrahydro phonium salts, methyl-, myristyl-, phenyl-, and benzyl furan, or a lower alcohol as a solution promoter. Also tri (C1-C4)alkyl- and cetryltrimethylammonium as well useful are perchloric acid or organic acids such as tri as (C1-C12)alkyl- and benzyltriphenylphosphonium 20 floroacetic, formic, and acetic acid, in combination with salts, with the compounds that have the larger and more catalytic amounts of mineral acids. The alkoxyalkox symmetrically structured cation generally proving to be ymethyl compounds in particular can also be cleaved by the more effective. using Lewis acids such as zinc bromide and titanium The introduction of the groups Ia, R5, and R15 by the tetrachloride in anhydrous medium, preferably in di procedures described above is generally carried out at a 25 reaction temperature between 0° C. and the boiling chloromethane or chloroform, with the 7-bromomethyl point of the particular reaction medium used, preferably or 7-bromozinc derivatives formed as intermediates between 20' and 130', optionally at elevated or reduced hydrolyzing spontaneously during the aqueous workup. pressure, for which the reaction time can amount to less In the cleavage in mineral acid solution, the reaction than 1 hour or up to several hours. 30 temperature must be chosen so that no significant dehy The reaction of the 3-alkylxanthines (VIII) to pro dration of the tertiary hydroxyalkyl group in the 1-posi duce the compounds pursuant to the invention of for mula OX) requires the introduction of two tertiary hy tion occurs; it should therefore be below 100 C. as a droxyalkyl groups. Either identical or different substitu rule. ents can be linked to the xanthine skeleton in succession, 35 The reduction of the xanthines of formulas (XI) and or two identical hydroxyalkyl groups can be linked (XV) with an oxoalkyl group in the position of Rs or without isolation of intermediates in a single-pot reac R1 to the corresponding hydroxyalkyl compounds can tion. indeed take place in principle either with base metals or The reductive cleavage of the benzyl and diphenyl methyl group from compounds of formula (XV) with by catalytic hydrogenation, but the method of choice the formation of the xanthine atom in the 7-position, is consists of the reaction occurring under the very mild carried out under standard conditions that were devel conditions and in high yields with simple metal hydrides oped especially in the framework of the protective (MH), complex metal hydrides (MMH)), or or group technique in alkaloid and peptide syntheses and ganometallic hydrides (Houben-Weyl, Vol. IV/l d can thus be assumed to be widely known. Besides the 45 (1981), pp. 267-282, and Vol. VI/1 b (1984), pp. chemical reduction, particularly of the benzyl com pounds with sodium in liquid ammonia (Houben-Weyl, 141-155). Of the numerous complex metal hydrides that Vol. XI/l (1957), pp. 974-975), the elimination of the can be used for the reduction of ketones, the most fre two aforementioned aralkyl groups by catalytic hydro quently used reagents might be mentioned, for example, genolysis using a precious metal catalyst is also espe 50 lithium alanate, lithium borohydride, and especially cially practical (Houben-Weyl, Vol. XI/l (1957), pp. sodium borohydride, that is easier to handle because of 968-971 and Vol. IV/Ic, Part I (1980), pp. 400-404). A its lower reactivity and above all permits working in lower alcohol is ordinarly used here as the reaction alcoholic, alcoholic aqueous, and pure aqueous solu medium (optionally with the addition of formic acid or tions or suspensions. In addition to the otherwise cus ammonia), or an aprotic solvent such as dimethylform 55 amide or particularly glacial acetic acid; however, their tomery inert solvents such as ethers (for example, di mixtures with water can also be used. Especially suit ethyl ether, tetrahydrofuran, 1,2-dimethoxyethane), able hydrogenation catalysts are palladium black and hydrocarbons and pyridine, nitriles such as acetonitrile palladium on activated charcoal or barium sulfate, can also be used as the reaction medium. The hydroge while other precious metals such as platinum, rhodium, nation, which is suitably carried out at temperatures and ruthenium frequently given rise to side reactions between O' C. and the boiling point of the particular because of competitive ring hydrogenation and are therefore only conditionally usable. The hydrogenoly solvent, but preferably at room temperature, generally sis is preferably carried out attemperatures between 20' occurs rapidly and is complete within several minutes to C. and 100 C. and at atmospheric pressure, or prefera 65 a few hours. bly slight excess pressure up to approximately 10 bar, The tertiary hydroxyalkylxanthines of formula (II) with reaction times of a few minutes to several hours can also be prepared by reacting substituted xanthines generally being needed. of formula CXIX) 5,272,153 19 20 OX) or (XI) or (XV), in which R2, R., R., R1, and n in (XIX) the formulas above have the meanings indicated above. R9 R10 The 3-alkylated mono- or dioxoalkyl- (XIXa), -(a)- haloalkyl) (XIXb), -(a)-alkoxycarbonylalkyl)- (XIXc), A. and -alkenylxanthines (XIXd) needed for this as starting CS materials are either known or can be prepared readily, ... l. for example, from the 3-alkyl-xanthines (VII) and the k sulfonyloxy- or haloketones (XVII) and (XVIII), co haloalkylsulfonates, or 1, co-dihaloalkanes (cf., for ex e) contain two identical or different groups of the 10 ample: V. B. Kalcheva et al., Journal fur prakt. Chemie formula 327 (1985) pp. 165-168), co-sulfonyloxy or co-halocar boxylic acid alkyl esters or sulfonyloxy or haloalkenes -(CH2)-CO-CH3 (XX); corresponding to formula (XXV) under the reaction Of conditions previously described in detail for the alkyla 15 tion of mono- and disubstituted xanthines with the con -(CH2)--CO-R' (XXI), pounds of formulas (VIII) and (Xa) In the organometallic reactions of the xanthines or only one substituent of the formula OXX) or (XXI), (XIXa) and (XIXc) functionalized in the R9 and R10 and hydrogen or the group R or R1 in the positions of 20 groups, the procedure is the same in principle as de R9 and R10, with (C1-C3)alkyl- or methylmetal com scribed for the preparation of the tertiary alcohols of pounds with reductive "alkylation' of the carbonyl formula (VIII) used as alkylating agents. Thus, the re groups to obtain the xanthines pursuant to the invention ductive alkylation of the ketones (XIXa) and of the of formulas (IX) to (XVI), or esters (XIXc) can take place, for example, with alkyl f) metallating xanthines of formula OXIX) that have potassium, -sodium, -lithium, -magnesium, -zinc, -cad two identical or different groups of the formula -(CH2 25 mium, -aluminum, and -tin compounds. The recently )-Hal (XVII), with Hal preferably standing for chlo recommended alkyltitanium and -zirconium compounds rine or bromine, or only one such group and hydrogen (D. Seebach et al., Agnew, Chem.95 (1983), pp. 12-26) or the substituent R or R15 in the other position, in the can also be used. However, since the alkylmetal com terminal position, and then reacting them with the ke pounds of sodium and potassium have a tendency tones of the formula 30 toward side reactions because of their high reactivity and those of zinc and cadmium are relatively sluggish, R-CO-CH3 (XVIII) the alkyllithium and -magnesium (Grignard) com with reductive alkylation of the carbonyl group to ob pounds are ordinarily preferred. tain the xanthines of formulas (IX) to (XVI) pursuant to 35 The strong nucleophilic organometallic compounds the invention, or are very sensitive to hydrolysis and oxidation. Their g) converting xanthines of formula (XIX) with the safe handling therefore requires working in anhydrous group medium, optionally under an inert gas atmosphere. The usual solvents or distributing agents are primarily those -(CH2)-COO-(C1-C4)alkyl (XXIV) 40 that are suitable also for the preparation of the alkyl metal compounds. Practical examples are especially in the positions of R9 and/or R10 and optionally hydro ethers with one or more ether oxygen atoms, for exam gen or the group R or R1 in the other position, by ple diethyl, dipropyl, dibutyl, or diisoamyl ether, 1,2- means of two equivalents of a methylmetal compound dimethoxyethane, tetrahydrofuran, dioxane, tetrahy per alkoxycarbonyl group, into xanthines of formulas 45 dropyran, furan, and anisole, and aliphatic or aromatic (IX) to (XVI) in which R" stands for methyl, or hydrocarbons such as petroleum ether, cyclohexane, h) converting xanthines of formula (XIX) having two benzene, toluene, xylenes, diethylbenzenes, and tetrahy identical or different groups of the formula dronaphthalene; however, tertiary amines such as tri ethylamine, or dipolar aprotic solvents such as hexame (XXV) SO thylphosphoric trianide, as well as mixtures of the sol -(CH2)--CH=CH vents mentioned can also be used successfully. The N reaction of the carbonyl compounds OXIXa) and (XIXc) CH3 with the Grignard compounds with the formula R MgHal can also beneficially be carried out by placing or only one such group and hydrogen or the group R. 55 the organometallic compound in an ether and adding or Rin the positions of R9 and R10, in which the group the ketone or the ester dropwise as a solution in dichlo (XXV) can contain the C=C double bond also in posi romethane or 1,2-dichloroethane. An addition of mag tion-isomeric arrangements on the branched carbon nesium bromide is frequently recommended, which is atom, for example, as -C=CH2, by acid-catalyzed able to increase the nucleophilicity of the organometal hydration obeying the Markownikoff Rule, into the lic compound because of its participation in the cornplex Xanthines of formulas (IX) to (XVI) pursuant to the cyclic transition state. invention, and if desired, then converting the tertiary The ketone or ester and the organometallic com hydroxyalkylxanthines of formulas Ib' and if obtained pound are generally combined at temperatures between pursuant to the invention by methods e) to h) that have -20' C. and 100 C., preferably between O' C. and 60', a hydrogen atom in the 1- or 7-position, optionally in 65 or at room temperature without external cooling, with the presence of basic media or in the form of their salts, the alkylmetal compound ordinarily being used in slight with the alkylating agents of formula (VIII) or (Xa) or excess. The reaction is then ordinarily completed by (XIII), into the trisubstituted compounds of formulas brief heating under reflux, for which times of several 5,272,153 21 22 minutes to a few hours are generally adequate. The Depending on the chain length of the alkyl group R' alkanolate formed is preferably decomposed with aque (at least C2) and/or the structure of a substituent R. (for ous ammonium chloride solution or dilute acetic acid. example, 2-hydroxypropyl), the tertiary hydroxyalklyx Metallic magnesium and lithium are primarily suit anthines of formula (II) can have one or two asymmet able for the metallation of the a-haloalkylxanthines ric carbon atoms and can thus be present in stereoiso (XIXb). On the other hand, the replacement of the meric forms. This invention therefore concerns both the halogen atom with lithium, which is also possible using pure stereoisomeric compounds and their mixtures. organolithium reagents, generally 1-butyl-, 2-butyl-, Effective amounts of the xanthines can be adminis t-butyl-, or phenyllithium, plays a subordinate role. tered to a subject by any one of various methods, for However, use is made especially of the Grignard com O example, orally as in capsule or tablets, or parenterally pounds, advantageously preparing them in the ethers, in the form of sterile solutions. The xanthines, while hydrocarbons, tertiary amines, or aprotic solvents listed effective themselves, can be formulated and adminis as particularly suitable for the reaction of the xanthines tered in the form of their pharmaceutically acceptable OXIXa) and (XIXc) with alkylmetal compounds, at addition salts for purposes of stability, convenience of temperatures between 25' and 125 C., preferably 15 crystallization, increased solubility, and the like. below 100 C. If the metallation reaction is carried out Preferred pharmaceutically acceptable addition salts in hydrocarbons, then the addition of an ether such as include salts of mineral acids, for example, hydrochloric tetrahydrofuran, or a tertiary amine such as triethylam acid, sulfuric acid, nitric acid, and the like; salts of ine in stoichiometric amount frequently proves useful. monobasic carboxylic acids, such as, for example, acetic The use of catalysts such as butanol, aluminum chloride, 20 acid, propionic acid, and the like; salts of dibasic car silicon tetrachloride, tetrachloromethane, and alumi boxylic acids, such as, maleic acid, fumaric acid, oxalic num or magnesium alcoholates may also be helpful. In acid, and the like; and salts of tribasic carboxylic acids, the halogen-metal exchange the chlorides ordinarily such as, carboxysuccinic acid, citric acid, and the like. react more slowly than the corresponding bromides and The xanthines can be administered orally, for exam iodides, but as a rule they provide better yields of or 25 ple, with an inert diluent or with an edible carrier. They ganometallic compound. To accelerate the beginning of can be enclosed in gelatin capsules or compressed into the reaction, the addition of some magnesium bromide, tablets. For the purpose of oral therapeutic administra some grains of iodine, or several drops of bromine, tion, the compounds can be incorporated with excipi tetrachloromethane, or methyl iodide with slight heat ents and used in the form of tablets, troches, capsules, ing is frequently recommended. The Grignard com 30 elixirs, suspensions, syrups, wafers, chewing gums, and pounds obtained are normally not isolated, but are re the like. These preparations should contain at least 0.5% acted immediately with the ketones of formula (XXIII) of active compound, but the amount can be varied de under the reaction conditions described for the reduc pending upon the particular form and can conveniently tive alkylation of the xanthines (XIXa) and (XIXc). be between 4.0% to about 70% of the weight of the The addition of water to the C-C double bond of the 35 unit. The amount of xanthine in such compositions is alkenylxanthines (XIXd) with the structural element of such that a suitable dosage will be obtained. Preferred formula (XXV), in which the hydroxy group adds to compositions and preparations according to the present the carbon atom with the fewer hydrogens to form invention are prepared so that an oral dosage unit form tertiary alcohols according to the Markownikoff Rule, contains between about 1.0 mgs and about 300 mgs of ordinarily occurs in aqueous solution or suspension in active compound. the presence of strong acids such as sulfuric, nitric, or Tablets, pills, capsules, troches, and the like can con phosphoric acid. Hydrogen halides and sulfonic acids tain the following ingredients: a binder, such as micro such as trifluoromethanesulfonic acid, acid exchange crystalline cellulose, gum tragacanth or gelatin; an ex resins, boron trifluoride complexes, or oxalic acid can cipient, such as starch or lactose; a disintegrating agent, also be used as catalysts. However, it is preferred to 45 such as alginic acid, Primogel, corn starch, and the like; operate in sulfuric acid, with an acid concentration of 50 a lubricant, such as magnesium stearate or Sterotes; a to 65% and temperatures of 0 to 10° C. being sufficient glidant, such as colloidal silicon dioxide; a sweetening as a rule. However, lower or higher acid concentration agent, such as sucrose or saccharin; or flavoring agent, and/or reaction temperatures can sometimes also be such as peppermint, methyl salicylate, or orange flavor used. In any case, the reaction temperatures should be SO ing. When the dosage unit form is a capsule, it can con kept as low as possible since the reverse dehydration to tain, in addition to material of the above type, a liquid the olefin can be disturbingly significant above approxi carrier, such as a fatty oil. mately 60' C. Other dosage unit forms can contain other materials The addition of a solvent inert to acids such as 1,4- that modify the physical form of the dosage unit, for dioxane, benzene, or toluene sometimes also provides 55 example, as coatings. Thus, tablets or pills can be coated benefits. Since esters can form as intermediates in the with sugar, shellac, or other enteric coating agents. A acid-catalyzed hydration, particularly when using the syrup may contain, in addition to the active compounds, high acid concentrations, it is recommended to treat the sucrose as a sweetening agent and preservatives, dyes, reaction batch with a large amount of water with brief colorings, and flavors. Materials used in preparing these heating after the action of the acid for the purpose of 60 compositions should be pharmaceutically pure and non ester hydrolysis, or to process the mixture in the alka toxic in the amounts used. line range. For purposes of parenteral therapeutic administra The experimental conditions for the optional conver tion, the xanthines can be incorporated into a solution sion of the 1- and 7H-compounds (IX) or (XVI) pursu or suspension. These preparations should contain at ant to the invention into the trisubstituted xanthines of 65 least 0.1% of the aforesaid compound, but may be var formulas (X) or (XI) or (XVI) by N-alkylation with the ied between 0.5% and about 50% of the weight thereof. compounds (VIII) or (Xa) of (XIII) have already been The amount of active compound in such compositions is described above in detail. such that a suitable dosage will be obtained. Preferred 5,272,153 23 24 compositions and preparations according to the present was purified by phenyl sepharose chromatography and invention are prepared so that a parenteral dosage unit FPLC to a final purity of greater than 99% as deter contains between 0.5 mg to 100 mgs of the active con mined by analysis on SDS acrylamide gels stained with pound. both Coomassie Brilliant Blue R250 and silver staining. Solutions or suspensions of the xanthines can also It has a molecular weight of 36,000 daltons by gel filtra include the following components: a sterile diluent, tion on Superose 12 (FPLC) and consists of 2 dimers of such as water for injection, saline solution, fixed oils, 17,000 daltons each. It was supplied sterile in phos polyethylene glycols, glycerine, propylene glycol or phate-buffered saline containing 0.1% bovine serum other synthetic solvents; antibacterial agents, such as albumin as a carrier protein (data supplied by Gen benzyl alcohol or methyl parabens; antioxidants, such as 10 ascorbic acid or sodium bisulfite; chelating agents, such zyme). Just before use, the rhi-TNF was diluted in as ethylenedianinetetraacetic acid; buffers, such as ace Hanks balanced salt solution containing 0.1% human tates, citrates or phosphates; and agents for the adjust serum albumin. ment of tonicity, such as sodium chloride or dextrose. The other materials were purchased as follows: Di The parenteral preparation can be enclosed in am 15 methyl sulfoxide (DMSO), n-formyl methionyl leucyl poules, disposable syringes or multiple dose vials made phenylalanine (FMLP; 10 mM stock solution in DMSO of glass or plastic. was stored in 20 l aliquots at -70' C.), heparin, cyto While dosage values will vary with the specific dis chrome c type VI from horse heart, and superoxide ease condition to be alleviated, good results are dismutase from bovine liver (SOD; stock solutions at 5 achieved when the xanthines of formula () or formula 20 mg/ml in Hawks balanced salt solution were stored in (II) are administered to a subject requiring such treat 100 ul aliquots at 70' C.) (Sigma Chemical, St. Louis, ment as an effective oral, parenteral or intravenous dose Mo.); Neutrophil isolation medium (NIM: Los Alamos or from 0.1 to 25 mg/kg of body weight per day. A Diagnostics, Inc., Los Alanos, N.M.); Hanks balanced particularly preferred effective amount is about 1.0 salt solution (HBSS), Minimum essential medium mg/kg of body weight per day. In general, daily dos 25 (MEM) and Medium 199 (M199) (Whittaker, M. A. ages will vary from 10-1,000 mg, preferably 100-600 Bioproducts, Walkersville, Md.); Dulbecco's phosphate mg per day. buffered saline (PBS; GIBCO Laboratories, Grand Is It is to be understood, however, that for any particu land, N.Y.); Limulus Amebocyte Lysate Test (LAL; lar subject, specific dosage regimens should be adjusted Associates of Cape Cod, Inc., Woods Hole, Mass); to the individual need and the professional judgment of 30 scrubbed nylon fiber (3 denier type 200) (Fenwal Labo the person administering or supervising the administra ratories, Deerfield, Ill.); Litex and Agarose type HSA tion of the xanthines. It is to be further understood that (Accurate Chemical and Scientific Corp., Hicksville, the dosages set forth herein are exemplary only and that N.Y.). they do not, to any extent, limit the scope or practice of PMN preparation: Purified PMN (-98% PMN and the invention. 35 >95% viable by trypan blue exclusion) containing

R6-C-(CH2) R4 65 wherein R is C1-C6 and p is 2, 3 or 4, the remain (CH)--CH, ing R or R3 being as defined above, and R2 is a OH C1-C4 alkyl group; 5,272,153 33 34 in which R stands for an alkyl group with 1 to 3 carbon atoms and n stands for a whole number from 2 to 5, the other R1 or R3 group that may optionally be present stands for a hydrogen atom *''' (II) or an aliphatic hydrocarbon group R with up to 6 5 N ) carbon atoms, whose carbon chain may be inter oes N N rupted by up to 2 oxygen atoms or may be substi tuted with a hydroxy or oxo group, or R2 b) an oxoallyl group of the formula 10 wherein at least one of R and R3 is either O a) a branched hydroxyalkyl group of the formula R-C-(CH2) R4 wherein R is C1-C6 and p is 2, 3 or 4, the remain 15 (CH)--CH, ing R or R being as defined above, and R2 is a OH C1-C4 alkyl group; wherein said amount is effective in inhibiting immune in which R stands for an alkyl group with 1 to 3 response by inhibiting the activity of IL-1, TNF, or carbon atoms and n stands for a whole number other leukocyte derived cytokines on monocytes. 20 from 2 to 5, the other R1 or R3 group that may 12. A method of inhibiting immune response in a optionally be present stands for a hydrogen atom mammal, wherein the method comprises administering or an aliphatic hydrocarbon group R with up to 6 to the mammal an amount of at least one xanthine of the carbon atoms, whose carbon chain may be inter formula: rupted by up to 2 oxygen atoms or may be substi 25 tuted with a hydroxy or oxo group, or R O R3 (II) b) an oxoallyl group of the formula O Y sA. 30 R6-C-(CH2) oes N N wherein R6 is C1-C6 and p is 2, 3 or 4, the remain R2 ing R1 or R3 being as defined above, and R2 is a C1-C4 alkyl group; wherein at least one of R and R3 is either 35 wherein said amount is effective in inhibiting immune a) a branched hydroxyalkyl group of the formula response by inhibiting the activity of IL-1, TNF, or other leukocyte derived cytokines on lymphocytes. R4 14. A method of alleviating an adverse condition in a mammal mediated by IL-1, TNF, or other leukocyte (CH)--CH, derived cytokines, which comprises administering to OH the mammal an amount of at least one xanthine of the formula: in which R stands for an alkyl group with 1 to 3 carbon atoms and n stands for a whole number from 2 to 5, the other R or R3 group that may 45 R1 O R3 (II) optionally be present stands for a hydrogen atom or an aliphatic hydrocarbon group R with up to 6 carbon atoms, whose carbon chain may be inter oss s rupted by up to 2 oxygen atoms or may be substi 50 tuted with a hydroxy or oxo group, or R2 b) an oxoallyl group of the formula wherein at least one of R1 and R3 is either O a) a branched hydroxyalkyl group of the formula 55 Ré-C-(CH2) R4 wherein R6 is C1-C6 and p is 2, 3 or 4, the remain (CH)--CH, ing R or R3 being as defined above, and R2 is a OH C1-C4 alkyl group; 60 u wherein said amount is effective in inhibiting immune in which R stands for an alkyl group with 1 to 3 response by inhibiting the activity of IL-1, TNF, or carbon atoms and n stands for a whole number other leukocyte derived cytokines on macro from 2 to 5, the other R1 or R3 group that may phages. optionally be present stands for a hydrogen atom 13. A method of inhibiting immune response in a 65 or an aliphatic hydrocarbon group R with up to 6 mammal, wherein the method comprises administering carbon atoms, whose carbon chain may be inter to the mammal an amount of at least one xanthine of the rupted by up to 2 oxygen atoms or may be substi formula: tuted with a hydroxy or oxo group, or 5,272,153 35 36 b) an oxoallyl group of the formula 17. A method of inhibiting immune response in a mammal, wherein the method comprises administering to the mammal an amount of at least one xanthine of the Ré-C-(CH), 5 formula: wherein R is C1-C6 and p is 2, 3 or 4, the remain ing R1 or R3 being as defined above, and R2 is a R O R3 (II) C1-C4 alkyl group; wherein said amount is effective in alleviating the 10 activity, which mediates the adverse condition, of Y s IL-1, TNF, or other leukocyte derived cytokines. oes N 15. A method of alleviating an adverse condition in a R2 mammal mediated by IL-1, TNF, or other leukocyte derived cytokines, which comprises administering to 15 wherein the mammal an amount of at least one xanthine of the formula: R and R2 and R3 are the same or different and are selected from the group consisting of O (I) (a) C1-C6 straight or branched chain alkyl; R. l 20 (b) C1-C6 straight or branched chain alkoxyalkyl; N N-a--ch, (c) C1-C6 straight or C1-C9 branched chain hy an 2 O droxyalkyl; O N (d) C1-C5 aliphatic hydrocarbon group whose car R2 25 bon chain may be interrupted by up to 2 oxygen atoms; in which (e) C3-C11 oxoalkyl group; R1 and R2 are the same or different and are indepen and in addition R1 and R2 can be cyclohexyl; dently selected from the group consisting of wherein said amount is effective in inhibiting immune straight-chain or branched alkyl radicals with 2 to 30 response by inhibiting the activity of IL-1, TNF, or 6 carbon atoms, cyclohexyl, straight-chain or branched chain alkoxyalkyl and hydroxyalkyl radi other leukocyte derived cytokines on neutrophils. cals; 18. A method of inhibiting immune response in a A represents a hydrocarbon radical with up to 4 mammal, wherein the method comprises administering carbon atoms, which can be substituted by a 35 to the mammal an amount of at least one xanthine of the methyl group; and formula: wherein said amount is effective in alleviating the activity, which mediates the adverse condition, of IL-1, TNF, or other leukocyte derived cytokines. *l JO orsR 16. A method of inhibiting immune response in a 40 N N mammal, wherein the method comprises administering to the mammal an amount of at least one xanthine of the ) formula: o? N N R2 (II) 45 wherein R1 and R2 and R3 are the same or different and are selected from the group consisting of 50 (a) C1-C6 straight or branched chain alkyl; (b) C1-C6 straight or branched chain alkoxyalkyl; (c) C1-C6 straight or C1-C9 branched chain hy wherein droxyalkyl; and in addition R1 and R2 can be R1 and R2 and R3 are the same or different and are cyclohexyl; selected from the group consisting of 55 (a) C1-C6 straight or branched chain alkyl; (d) C1-C6 aliphatic hydrocarbon group whose car (b) C1-C6 straight or branched chain alkoxyalkyl; bon chain may be interrupted by up to 2 oxygen (c) C1-C6 straight or C1-C9 branched chain hy atons; droxyalkyl; (e) C3-C11 oxoalkyl group; (d) C1-C6 aliphatic hydrocarbon group whose car wherein said amount is effective in inhibiting immune bon chain may be interrupted by up to 2 oxygen response by inhibiting the activity of IL-1, TNF, or atons; (e) C3-C11 oxoalkyl group; other leukocyte derived cytokines on mononuclear and in addition R1 and R2 can be cyclohexyl; phagocytes. wherein said amount is effective in inhibiting immune 65 19. A method of inhibiting immune response in a response by inhibiting the activity of IL-1, TNF, or mammal, wherein the method comprises administering other leukocyte derived cytokines on polymorpho to the mammal an amount of at least one 7-(oxoalkyl) nuclear leukocytes. 1,3-dialkyl xanthine of the formula: 5,272,153 37 38 R1 and R2 and R3 are the same or different and are O R3 (II) selected from the group consisting of R / (a) C1-C5 straight or branched chain alkyl; N N (b) C1-C6 straight or branched chain alkoxyalkyl; (c) C1-C6 straight or C1-C9 branched chain hy o? OC)N N 5 droxyalkyl; (d) C1-C6 aliphatic hydrocarbon group whose car k bon chain may be interrupted by up to 2 oxygen atoms; wherein O (e) C3-C1 oxoalkyl group; R1 and R2 and R3 are the same or different and are and in addition R1 and R2 can be cyclohexyl; selected from the group consisting of wherein said amount is effective in inhibiting immune (a) C1-C6 straight or branched chain alkyl; response by inhibiting the activity of IL-1, TNF, or (b) C1-C6 straight or branched chain alkoxyalkyl; otherleukocyte derived cytokines on lymphocytes. (c) C1-C6 straight or C1-C9 branched chain hy 15 22. A method of alleviating an adverse condition in a droxyalkyl; mammal mediated by IL-1, TNF, or other leukocyte (d) C1-C5 aliphatic hydrocarbon group whose car derived cytokines, which comprises administering to bon chain may be interrupted by up to 2 oxygen the mammal an amount of at least one xanthine of the atoms; formula: (e) C3-C11 oxoalkyl group; 20 and in addition R1 and R2 can be cyclohexyl; wherein said amount is effective in inhibiting immune (II) response by inhibiting the activity of IL-1, TNF, or R r other leukocyte derived cytokines on monocytes. 20. A method of inhibiting immune response in a 25 N ly mammal, wherein the method comprises administering oals N s' to the mammal an amount of at least one xanthine of the formula: 30 wherein (II) R and R2 and R3 are the same or different and are selected from the group consisting of (a) C1-C6 straight or branched chain alkyl; (b) C1-C6 straight or branched chain alkoxyalkyl; (c) C1-C6 straight or C1-C9 branched chain hy 35 droxyalkyl; (d) C1-C6 aliphatic hydrocarbon group whose car bon chain may be interrupted by up to 2 oxygen wherein atoms; R and R2 and R3 are the same or different and are (e) C3-C1 oxoalkyl group; selected from the group consisting of and in addition R1 and R2 can be cyclohexyl; (a) C1-C6 straight or branched chain alkyl; wherein said amount is effective in alleviating the (b) C1-C6 straight or branched chain alkoxyalkyl; activity, which mediates the adverse condition, of (c) C1-C6 straight or C1-C9 branched chain hy IL-1, TNF, or other leukocyte derived cytokines. droxyalkyl; 45 23. The method as claimed in any one of claims (d) C1-C6 aliphatic hydrocarbon group whose car 16-22, wherein R1 is a hydroxyalkyl radical. bon chain may be interrupted by up to 2 oxygen 24. The method as claimed in claim 23, wherein R2 is atoms; a straight-chain or branched alkyl radical with 1 to 6 (e) C3-C11 oxoalkyl group; carbon atoms. and in addition R1 and R2 can be cyclohexyl; 50 25. The method as claimed in claim 24, wherein the wherein said amount is effective in inhibiting immune R3 group stands for an aliphatic hydrocarbon group R5 response by inhibiting the activity of IL-1, TNF, or with up to 6 carbon atoms. other leukocyte derived cytokines on macro 26. A method of inhibiting immune response in a phages. mammal, wherein the method comprises administering 21. A method of inhibiting immune response in a 55 to the mammal an amount of at least one xanthine of the mammal, wherein the method comprises administering formula: to the mammal an amount of at least one xanthine of the formula: O R3 (II) R / O R3 (II) 60 N N R / ) N N ) o? N N O 2 N N k 65 R2 wherein R and R2 and R3 are the same or different and are wherein selected from the group consisting of 5,272,153 39 40 (a) C1-C6 straight or branched chain alkyl; (b) C1-C6 straight or branched chain alkoxyalkyl; O R3 (II) R1 / (c) C1-C6 straight or C1-C9 branched chain hy N N droxyalkyl; N and in addition R and R2 can be cyclohexyl; wherein said amount is effective in inhibiting immune o? N C)N response by inhibiting the activity of IL-1, TNF, or k other leukocyte derived cytokines on polymorpho nuclear leukocytes. 10 wherein 27. A method of inhibiting immune response in a R1 and R2 and R3 are the same or different and are mammal, wherein the method comprises administering selected from the group consisting of to the mammal an amount of at least one xanthine of the (a) C1-C6 straight or branched chain alkyl; formula: (b) C1-C5 straight or branched chain alkoxyalkyl; 15 (c) C1-C6 straight or C1-C9 branched chain hydroxy alkyl; r O / R3 (II) and in addition R1 and R2 can be cyclohexyl; N wherein said amount is effective in inhibiting immune N response by inhibiting the activity of IL-1, TNF, or 20 other leukocyte derived cytokines on monocytes. O OC)N N 30. A method of inhibiting immune response in a k mammal, wherein the method comprises administering to the mammal an amount of at least one xanthine of the wherein 25 formula: R and R2 and R3 are the same or different and are selected from the group consisting of R O R3 (II) (a) C1-C6 straight or branched chain alkyl; (b) C1-C6 straight or branched chain alkoxyalkyl; (c) C1-C6 straight or C1-C9 branched chain hy 30 droxyalkyl; l, l? and in addition R1 and R2 can be cyclohexyl; wherein said amount is effective in inhibiting immune response by inhibiting the activity of IL-1, TNF, or 35 wherein other leukocyte derived cytokines on neutrophils. R and R2 and R3 are the same or different and are 28. A method of inhibiting immune response in a selected from the group consisting of mammal, wherein the method comprises administering (a) C1-C6 straight or branched chain alkyl; to the mammal an amount of at least one xanthine of the (b) C1-C6 straight or branched chain alkoxyalkyl; (c) C1-C6 straight or C1-C9 branched chain hy formula: droxyalkyl; and in addition R and R2 can be cyclohexyl; R O Rr wherein said amount is effective in inhibiting immune N N response by inhibiting the activity of IL-1, TNF, or N 45 other leukocyte derived cytokines on macro phages. o? N C)N 31. A method of inhibiting immune response in a R2 mammal, wherein the method comprises administering 50 to the mammal an amount of at least one xanthine of the wherein formula: R and R2 and R3 are the same or different and are selected from the group consisting of R1 O R3 (II) (a) C1-C6 straight or branched chain alkyl; (b) C1-C6 straight or branched chain alkoxyalkyl; 55 Ys s (c) C1-C6 straight or C1-C9 branched chain hy droxyalkyl; l? and in addition R1 and R2 can be cyclohexyl; wherein said amount is effective in inhibiting immune response by inhibiting the activity of IL-1, TNF, or wherein R1 and R2 and R3 are the same or different and are other leukocyte derived cytokines on mononuclear selected from the group consisting of phagocytes. (a) C1-C6 straight or branched chain alkyl; 29. A method of inhibiting immune response in a 65 (b) C1-C6 straight or branched chain alkoxyalkyl; mammal, wherein the method comprises administering (c) C1-C6 straight or C1-Cs branched chain hy to the mammal an amount of at least one 7-(oxoalkyl) droxyalkyl; 1,3-dialkyl xanthine of the formula: and in addition R1 and R2 can be cyclohexyl; 5,272,153 41 42 wherein said amount is effective in inhibiting immune Ri and R2 and R3 are the same or different and are response by inhibiting the activity of IL-1, TNF, or selected from the group consisting of other leukocyte derived cytokines on lymphocytes. (a) C1-C6 straight or branched chain alkyl; 32. A method of alleviating an adverse condition in a (b) C1-C6 straight or branched chain alkoxyalkyl; mammal mediated by IL-1, TNF, or other leukocyte 5 (c) C1-C6 straight or C1-C9 branched chain hy derived cytokines, which comprises administering to droxyalkyl; and in addition R1 and R2 can be cyclohexyl; the mammal an amount of at least one xanthine of the wherein said amount is effective in alleviating the formula: activity, which mediates the adverse condition, of O IL-1, TNF, or other leukocyte derived cytokines. (II) 33. The method as claimed in any one of claims 26-32, wherein R1 is a hydroxyalkyl radical. 34. The method as claimed in claim 33, wherein R2 is a straight-chain or branched alkyl radical with 1 to 6 15 carbon atoms. 35. The method as claimed in claim 34, wherein the R3 group stands for an aliphatic hydrocarbon group Rs with up to 6 carbon atoms. wherein 20

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