CA2039458A1 - Methods for treating interleukin-1 mediated diseases - Google Patents

Abstract

Abstract of the Invention Disclosed are medicines for treating interleukin-1 mediated diseases including interleukin-1 mediated arthritis, interleukin-1 mediated inflammatory bowel disease, interleukin-1 mediated septic shock, interleukin-1 mediated ischemia injury, and interleukin-1 mediated reperfusion injury. The medicine com-prises a therapeutically effective amount of an interleukin-1 inhibitor. In a preferred embodiment, the interleukin inhibitor is selected from the group consisting of IL-lraa, IL-lraB, IL-lrax, and a combination thereof. A preferred method of produc-ing IL-lraa, IL-lraB, Il-lrax is by recombinant DNA technology.

Description

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B~CKGROUND OF THE INVENTION
~ he present invention describes drugs for the treatment of a variety of diseasas and medical conditions.
The common elem~nt of the diseases and medical conditions that axe suitable ~or treatment according to the methods described her~in is an involvement o~ interleukin-1. This invention describes methods for the treatment of interleukin-l mediated diseases and medical conditions.
Cytokines are extracellular proteins which modify the behavior o~ cells, particularly those cells that are in the immediate area of cytokin2 synthesi~ and release. one of the most potent inflammatory cytokines yet discovered and a cytokine which is thought to be a key mediator in many diseases and medical conditions is interleukin-l (IL-1).
Interleukin-l, which is manufactured, though not exclusively, by cells of the macrophagaJmonocyte lineage, may be produced in two forms, IL-l alpha (Il-la) and IL 1 beta (IL-lb).
A disease or madical condition is considered to be a "interleukin-l mediated disease" i~ the spontaneous or experimental disease or ~edic~l condition is as~ociated with elevated level~ of IL-l in bodily fluid~ or tissua or if cell~ or tis~ues taken fro~ the body produce elevated levels of IL 1 in culture. In ~any cases, such interleukin~1 mediated diseasss are also recoyniz2d by he following additional two conditions~ pathological findings associated wlth th~ diseasQ or medical condition can be mimicked experimentally in animal~ by tha admini~tration of IL-l; and (~ the pathology induced in experimental animal models of the clisease or medical condition can be inhibited or abolished by treatment with agents which inhibit the action of IL-1~ In most "Interleukin-1 mediated diseases'~

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at least two of the three conditions re. met, and in many "interleuXin-1 mediated diseases" all three conditions are met. A list of diseases or medi.cal conditions which are interleukin~l mediated includes, but i5 no~ limi~ed to, the following:
1) Arthritis 2) Inflammatory Bowel Disease 3) Septic 9,hock 4) Ischemic: injury 5) Reperfusion injury 6) Osteoporosis 7~ Asthma 8) Insulin diabetes 9) ~yelogenous and other leukemias 10) Psoriasis 11) Cachexia/anorexia Arthritis is a chronic joint ~isease which afflicts and disable~, to varying degrees, millions of people worldwide. The disease is typically characterized at the microscopic lev~l by the inflammation o~ synovial tissue and by a progressive degradation of the molecular components constituting the ~oint cartilage and bone. Continued inflammation and erosion of the joint frequently lead to con iderable pain, swellin~, and loss of ~unction.
~ hile the etiology of arthritis is poorly understoodt considerable information has recently been gained regarding the molecular aspects of inflammation.
This research has lead to the identi~ication o~ certain cytokines, which are believed to figure prominently in the mediation of in~lammation. The involYement o~ interleukin-l in arthritis has been implicated by two distinct lines of evidence. First! increased levels o~ interleukin-1 and of the mRNA encoding it have been ~ound in the synovial tissue and nuid of arthritic join~s. Re~erencas of interest include G. Buchan e~ ~1., Third Annual ~eneral Meeting of -2=

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the ~ritish Society for Rheumatology, London, England, November 19-?1, 1988, PR. J. Rhieumatol 25 (Supplemental 2) 1986; A. Fontana et al., Rheumatology Int., 2, pp. 49~53 (1982); and G. Duff et al, Mono:kines and Oth~r Non-Lymphocytic Cytokines, M. Powanda et al., editors, pp. 387 392, 1988 Alan R. Liss, Inc.
Second, the administration o~ interleukin~1 to healthy joint tissue has been shown on numerous sccasions to result in the erosion of cartilage and bone. In one experiment, intraarticular injections oP I~ 1 into rabbits were ~hown to cause cartilage destruction ln vivo as described by E. Pettipher ~_3~., Proc. Natl. Acad. Sci.
USA, Vol. 83, pp. 8749-8753, ~ovember, 1986. In other s udies, IL-l was shown to cause the degradation of both cartilage and bone in tissue explants. Relevant references include J. SaXlatavala ~ lo~ Development of Diseases of Cartilage and Bone Matrix, Alan R. Liss, Inc.~ pp. 291~298, and P. Stashenko et al., The American Association of I~munologists, Vol. 138, pp. 1464-1468, No. 5, March 1, 1987.
One generally accep~ed theory used to explain the causal link between IL-l and inflammation is that IL-l sti~ulates various cell types, such as ~ibroblasts and chondrocytes, to produce and secrete proinflammatory or degradative compounds, such as prostaglandin E2 and collagenase Consequently, the present inventors postulated that substanc that inter~ere with the aetivity o~
interleukin-l would appear to make excellent candidates ~or use in the treatment o~ inflammatory diseases like arthritis.
Inflammatory bowel disease ("IBD") is a term used to describe both acute and chronic in~lamma ory conditions of the tissue of th~ intestinal tract and encompasses two generally distinct maladies known as Ulcerative Colitis and Crohn's disease. Ulcerative Colitis is a mucosal ulceration of the colon. Crohn's disease, which is also referred to as Ileitis, Ileocolitis and Colitis, is a transmural inflammation that can be found throughout the yeneral intestinal tract.
IBD is characterized by various histological features including transmural acute and chronic granulomatous inflammation with ulceration, crypt abbesses and marked fibrosis. Not all of these indications will be found in all IBD cases. Spontaneous reactivation, extraintestinal inflammation and anemia are o~ten associated with IBD. Large joint arthritis is commonly found in pati2nts suffering from Crohn's disease.
A5 is found in the molecular processes of the in~lammation associated with arthritis, research has found that various cytokines appear to mediate aspects of IBD. In particular, IL-l has been implicated as a mediating material in IBD. Again, two distinct lines of evidence lead to this conclusion.
Increased levels of IL-l have been found in aff2cted areas of intestines fro~ patients with I8D.
Tissues from pati~nts with active Ulcerative Colitis showed IL-1 levels about 15 times the level found in control samples. TissuQs with active Crohn's disease showed IL-l levels about 6 times that of ths control, and tissues with inac~ive Crohn's diseasa were about 3 times that o~ the control tissue sa~ples. Sartor çt ~1. Gastroenterology, 94, Pg. A399)(Abstract o~ Paper). See also 5a sangi et al.
Clin. Exp. Immunol., 67, Pp. 594 605 ~1987); Rachmilewitz et al. Gastroenterology, 97, Pg. 326 (19B9) (the bioassay used th~rein to determine IL-1 concentration levels is known to also unselectively detect lL-2, IL-4, IL-6 and IL-7~.
Th2 role of IL-l in IBD ~a~ also been implicated by studies that have shown that the perfusion of rabbit colons with IL-1 induces the production of prosta~landin and thromboxane. Comminelli ~ 31- Gastroenterology, 97, Pp.

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1400-1405 (1989). This is consistent w:ith the hypotheSis ~escribed above, that IL-l is linked to the in~lammation of tissues due to its stimulation of the production of proinflamm~tory or degradativa compounds. Thus, it is likely that systemic and local IL-l production initiates or contributes to the infla~matory response in IBD, and plays an active role in the pathogenesis of the disease. The systemic production of IL-l may also be responsible, in part, for the extraintestinal inflammation associated with Crohn's disease.
These re~ults have led the inventors hereo~ to propose that substances that would interfere with the activity of interleukin-1 could be ef~ective compounds for the treatment of IBD.
Septic shock is a condition as~ociated with massive bacterial invasion. It is commonly believed that the shock is brought on, at least in part, by the presence o~ bacterial toxins (e.g., liyopolysaccharides). Septic shock is a relatively common causa of mortality in the hospital setting. At present there are few treatment Qpti.ons for patients sufferi.ng from septic shocX~ a~d the treatments available are generally supportive in nature.
5eptic shock is characteri2ed by various symptoms including a drop in ~ean art~rial blocd pressure tMAP), a decrease in cardiac output, tachycardia, tachypnea, lacticacidemia and leukopenia. Yarious ~ytokines, including interleukin-l, hava been implicated in the mediation of septic shock, although the specific etiolo~y of the disease is not ~ully understood.
That IL-l may have a role in the mediation of septic shock has been suggested by two lines o~ evidence.
one study has been conducted wherein the blood serum of children suffering from gram-negative septicemia was analyzed for IL-l concentration. Thi~ study showed that elevated levP~ls of IL-1 were found in 21% o~ the patients 2 ~

~xamin~d. In addition, it was shown that IL-l serum levels were significantly higher in patients who died than in the survivors. Girardin ~t_al. New England J. of M~dicine 319, pp~ 397-400 (1988). See alsQ, Cannon et al. Critical Care Medicine, p. S58 (Abstract) April, 1989 (increased I~-l levels in patients suffering ~rom sepsis ~yndrome).
It has also been shown that human IL-l induces shock-like state in rabbits. A single bolus injection o~
human IL-lb brought about hypotension and several hemodynamic and hematological parameters characteristic o~
septic shock. For example, the M~P of IL-1 injected ra~bits decrea~ed by a minimum of 19.1%. Okusawa et al., J. Clin.
Invest., 81, pp. 1162-1171 (198~).
These result~ hav~ led the inventors hereof to propose that substanc~s which interfere with the activity of interleuXin-l could be effective compounds for the treatment of septic shock.
Ischemic injury may occur to a tissue or organ whenever that tissue or organ is depriv~d of its normal blood flow. Further damage may occur wh~n the flow of oxyg~nated blood is restored to that tissue. The extent and reversibility of th~ damage imparted depends, partly, on the severity o~ the original insult. It is possible, however, to mitigate the ~xtent o~ tissue damage resulting from reperfusion by a variety of th~rapeutic interventions.
Simpson PJ ~t al. In: ~alliw~ll B. (ed) Ox~q~L_adicals an~
Tlssu~ Intury, Brook ~odge Symp - Upjohn (1988).
Reperfusion injury is a well documented sequela to ische~ic episodes in thQ h~art, gut, kidney, liver and other organs. S~mpson PJ ~ . In: Halliwell B~ (ed) Oxyaen Rad~als and Tissu~ Iniuxy, Brook Lodge S~mp - Upjohn (1988); H~rman B. et ~1. FASEB J. 2:1460151 (1988); McDougal WS. Th~ J. o~ Urology, 1~0:1325-1330 (1988); Finn WF. Kidney Int., 7:171-182 (1990); Schrier XW. Klin Wochenschr, 66:800-807 (lg88); and Winchel RJ. Transplantation 48:393-3g6 (1989). The exact pathogenesis of reperfusion injury may vary depending on the tissue aPfected. In the heart, for instance, reperfusion injury is accompanied by a dramatic influx of neutrophils, and these cell5 are thought to play a major role affecting the reperfu.sion damage. Lucchesi BR
et al. Ann Rev Pharmacol Toxicol 26:201-224 (1988). Renal ischemia and reperfusion injury, on the other hand, appear to involve an increase in tubula,r cell membrane permeability, increa~ed levPls of intracellular calcium, altersd mitochondrial re~pira~or~ function, and the generation of ~ree radicals. In the kidney, the role of extravasating neutrophils in affecting the reperfusion injury is less certain. McDougal WS. The J. Urology, 140:1325 1330 ~19883; Finn WF. Xidney Int., 37:171-182 ~1990); Schrier RW. Rlin ~ochenschr, 66:800~807 (1988); and Winchel RJ. Transplantation 48:393-3~6 (1989).
Despite the differences in csllular participa~ion during ischemia and reperfusion inju~y, there may he similarities in the und~rlying mechanism. Interleukin-1 is recognized as an early stage mediator of organ injury, and may be generated by re~ident or newly infiltrated in1ammatory cells giving rise to organ specific tissue pathology. In such an in~tance, the ability to inhibit the biological activity of IL-l would represent a novel therapeutic intervention aimed at limiting the extent of tissue damage.
This information has led the inventors hereof to propo~e that substances which interfere with the activity of interleukin-l could be e~ective compounds for the minimization of ischemia and reperfu~ion injury.
To date, an ef~ective, yet ~elective, inhibitor of IL-l has not been available in su~ficient quantities or purity to prove that IL 1 is a target for pharmaceutical intervention in the treatment o~ arthriti~ D, septic shock, ischemia injury or reperfusion injury and for use as a therapeutic agent in the treatment of inflammation.
Despite the prior art, the present inventors have iden-tified a class of compounds, referred to herein as interleukin-l inhibitors, that prevent and treat interleukin-l mediated diseases.
Moreover the present therapeutic interventions may be practiced without unacceptable compromise of normal physiological processes (e.g. immuno competency) which are essential to the patient'swell being.
In currently pending United States Patent Application Serial No. 07/506,522, filed April 6, 1990 specifically incor-porated herein by reference, a preferred class of naturally occurring, proteinaceous interleukin-l inhibitors and a method for manufacturing a substantial quantity of the same with a high degree of purity are described. In particular, the aforementioned application describes in detail three such interleukin-l inhibitors which are interleukin-l receptor antagonists (IL-lra's), namely, IL-lialpha (IL-lia), IL-libeta IL-liB, and IL-lix. These IL-lra's will be referred to as IL-lraa, IL-lrab and IL-lrax, respectively, in this application.
SUM~lARY OF THE INVENTION
The present invention describes a medicine (or a therapeutic agent) for the prevention or treatment of interleukin-1 mediated diseases of patientsin need thereof.
An object of the present invention is to provide a therapeutic agent for treating interleukin-l mediated diseases, such as interleukin-l mediated arthritis, interleukin-l mediated 2~39~

inflammatory bowel disease ("IBD"), interleukin-l mediated septic shock, interleukin-l mediated ischemia injury and interleukin-l mediated reperfusion injury.
Additional objects and advantages of this invention will be set forth in part in the description which follows and in part will be obvious from the description or may be learned from practice of the invention. The objects and advantages may be realized and attained by means of the instrumentalities and com-binations particularly pointed out in the appended claims.
To achieve the objects and in accordance with the purposes of the present invention, medicines are disclosed for treating interleukin-l mediated diseases, including interleukin-l mediated arthritis, interleukin-l mediated inflammatory bowel disease, interleukin-l mediated septic shock, interleukin-l mediated ischemia injury and interleukin-l mediated reperfusion injury. These medicines comprise a therapeutically effective amount of an interleukin-l inhibitor in admixture with a pharma-ceutically acceptable carrier. The medicines are practically almost always placed in packages for commercial use. The packages may carry indications or instructions that the medicines can be used for preventing or treating such interleukin-l mediated diseases.
Preferred interleukin-l inhibitors of the present invention are proteins and, more particularly, are naturally-occurring proteins. The naturally-occurring proteins are preferred because they pose cL relatively low risk of producing unforeseen ":
-side effects in patients treated therewith.
A preferred class of interleukin-l inhibitors are the human proteins which naturally serve as interleukin-l recep-tor antagonists (IL-lra's). Preferably, those IL-lra's which are preferred in the practice of the present invention are selected from the group consisting of IL-lraa, IL-lraB, IL-lrax, or the N-terminal methionyl derivatives of IL-lrax. Also pre-ferred are proteins which have been modified for example by the addition of polyethylene glycol (PEG) or any other repeat polymer to increase their circulating half-life and/or to decrease their immunogenicity.
While the production of IL-lra may be achieved by extraction from naturally available sources, such as by isolation from the conditioned medium of cultured human - 9a -:,:

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monocytes, a preferred method of IL-lra production is by recombinant DNA technology. Recombinant DNA technology is prsferred in part because it is capable of producing comparativPly high~r amounts of IL~lra at greater purities.
It is to be understood that both the foregoing general description and the ~ollowing detailed description are exemplary and explanatory only and are not restrictive of the invantion as claimed.

BRIEF DESCRIPTION pF T~E DRAWINGS
FIG. 1 depicts the release of glycosaminoglycans (GA~) from bovine nasal cartilage in rssponse to increasing amounts of IL-lB;
FIG. 2 depicts the inhibitory ef~ect of increasing amounts of IL-lra on the IL-lB-induced release of ÇAG from bovine nasal car~ilage;
FIGS. 3 and 4 depict the inhibitory effect of IL-lra on the pathogenesis of type II collagen~induced arthritis in mice:
FIG. 5 depicts the inhibitory eff2ct of IL-lra on SCW reactivation of SCW-induced arthritis in rats.
FIG. 6 depicts the effects of IL-lra treatment on various indications of interleukin-l mediated IBD, FIG. 7 depicts the e~fects of IL-lra on PG-APS
reactiYation of joint in~lammation in conjl~nction with indomethacin.
FIG. 8 depicts the effects o~ IL-lra on survival rate in rabbits with Pndotoxin~induced shock.

ETAILED DESCRIPTION OF_PREFERRED EMBODIMENTS
Re~erence will now be made in detail to the presPntly preferred embodiments of the invention, which, together with ~he following examples, serYe to explain the principles of the invention.
As noted aboYe, the present invention relates to 2~3~

medicines for treating in-terleukin-l mediated di.s~ases including in~erleukin-1 mediated arthritis, interleukin-l mediated inflammatory bowel disease, interleukin-l mediated septic shock, interleukin-l mediated ischemia injury, and interleukin-l mediated reperfusion injury in patients sufferinq therefrom. This medicine co~prises a therapeutically effective amount of an interleukin-l inhibitor. In one Qmbodimsnt, th0 preferred interleukin-l inhibitors of the present invention are naturally-occurring proteins that serve as IL~1 receptor antagonists (IL-lra's).
A disease or medical condition i~ considered to be a 1l interleukin-l mediated disease" if the spontaneous or experimental disease or medical condition is associated with elevated levels of IL-l in bodily fluid3 or tissue or if cellc or tissues taken from the body produca elevated levels of IL-l in culture. In many cases, such in erleukin-l mediated diseases are also recognized by the following additional two conditions: (1) pathological findings associated with the diseas~ or m~dical condition can b~
mimicked experimentally in animal 5 by the administration oP
IL-1: and (2) th~ pathology inducQd in experimental animal modsls of the diseasQ or medical condition can be inhibited or abolished by treatment with agents which inhibit the action o~ IL-l. In mo3t "Interleukin-1 ~ediated diseases"
at least two of the three condition~ are met, and in many "interleukin-l mediated diseases" all thre~ conditions are met. A list of disea-e~ or medical conditions which are int~rleukin-l mediated includa~, but is not limited to, the following:
1) Arthritis 2) Inflammatory Bowel ~isease 3) Septic 5hock 4) Ischemic injury 5) Reparfu~ion injury 2~4~g 6) Osteoporosis 7) A thma 8) Insulin cliabetes 9) Myelogenous and other leukemias 10) Psoriasis;
11) Cachexia/anorexia The naturally-occurrinq protein6 are preferred in part because th~y pose a comparat:ively low risk of produoing un~oreseen and undesirabl9 physiological sid~ efgects in patients treated therewith.
For purpose~ oX ths specification and claims, a protein is deemed to be I'natural:Ly-occurring" if it or a sub~tantially equivalent protein can b~ found to exist normally in healthy humans. "Naturally occurring" proteins may b~ obtainQd by r~combinant DNA methods as well a~ by isola.ion fro~ cells which 3rdinarily produce them.
"Naturally-occurring" encompasse~ prot~in~ that contain an N terminal methionyl group as a consequenc~ of expression in . Coli.
"Substantially equivalent" as used throu~hout the specification and claim~ is defined to mean possessing a VQry high degree of a~ino acid residue homology (See qene~ally ~. Dayho~ s.og Protei~ Seq~ence and ~t~UC~U~, Vol. 5, p. 124 (1972), Na~ional Biochemical Research Foundation, Wa~hington, D.C., SpQ ifically incorporated herein by re~rence~ as w~ll as possessing comparable ~iological a¢tivity.
~ articularly pre~erred IL-lra's o~ ~he present invention arQ th~ naturally-ocGurring proteins that exist n YiV~ a~ regulator~ o~ interleukin-l that have previously been described in a currcntly pending Un~ted States patent application. This application is U.~. Patent Application SPrial No. 07~266,531, ~ d November 3, 198B, by Hannum e~
-, which i5 entitled "Interleukin-l Inhibitors."

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Three preferred forms of IL-lra, each being derived from th~ same DNA coding sequence, were disclosed and described in the aforementioned Hannum et al.
application. The first of these, IL-lraa, was characterizsd as behaving as a 22-23 kD molecule on SDS-PAGE with an approximate isoelectric point of 4.8, eluting from a Mono Q
FPLC column at around 52 mM NaC1 in Tris buffer, p~ 7.6.
The second, IL-lraB, was charact~rized as behaving as a 22-23 kD protein, p.I=4.8, eluting from a ~ono Q column at 60 mM NaC1. The third, IL-lrax, was characterized as behaving as a 20 kD protein, eluting from a Mono Q column at 48 mM
NaCl. All thrPe o~ the Hannum ~t al. interleukin 1 inhibitors wer~ shown to possess similar functional and immunological activities. The present inventlon also includ2s modified IL-lra's. In one e~bodiment, the IL-lra is modified by attachment of one or more polyethylene glycol (PEG) or other repeating polymeric moietie~. In another embodiment, the IL-lra contain~ an N-terminal methionyl group as a consequence of expression in E~ coli.
Methods for producing the Hannum et al. inhibitor~
are also disclosed in th~ above mentioned application. One disclosed method consisted of isolating the inhibitors ~rom human monocyt~s (where thPy ars naturally produced). A
second disclosed method involved isolating the gene responsible for coding the inhibitors, cloning the gene in suitable vector~ and cell types, expressing the g~ne to producQ tha inhibitors and harvesting the inhibitors. The latter ~ethod, which is exemplary o~ recombinant DNA m~thods in gen ral, is a preferred method o~ tha present inYention.
Recombinant DNA method~ are preferr~d in part becausP they ~re capable o~ achieving comparatively higher amounts at greater purities.
Additional interleukin 1 inhibitors includa compounds capable o~ specifically preventing activ~tion of 2~3~

cellular receptors to IL-1, Such compounds include IL-1 binding prote.ins such as soluble r~ceptors and monoclonal antibodies. Such compounds also include receptor antagonists and monoclonal antibodi~s to the receptors.
A second class of IL-lra's include the compounds and proteins which ~lock in vivc! synthesis and/or extracellular release of IL-l. Such compounds include agents which af ct transcription of IL-l genes or processing IL-1 preproteins. Under certain conditions, the IL-lra will block IL-l induced IL-1 production.
Preferably, the above de~cribed IL lrals are produced by the aforementioned method in "substantially pure" form. By "substantially pure" it is meant that th~
inhibitor, in an unmodified foxm, has a comparatively high specific activity, preferably in the range of approximately 150,000-500,000 receptor units/mg as defined by Hannum et al. in Nature 34~: 336-340 (1990) and Eisenberg et al. in Nature 343~ 341-346 (1390), both of which are specifically incorporated herein by reference. It is to be recogniz~d, how~ver, that derivatives of IL-lra may have different specific activities. In a preferred embodiment of the present invention, a therapeutic co~position comprising at least one o~ IL lraa, IL-lraB, and IL-lrax is administered in an effective amount to patients suf~ering ~rom interleukin-1 mediated diseases.
Because it is possible that the inhibitory function of the preferred inhibitors is imparted by one or mor~ discrete and separable portions, it is al50 envisioned that the method of the pre~ent invention could be practiced by ad~inistering a therapeutic composition whose active ingredient consists o~ that portion (or those portions) o~
an inhibitor which controls (or control) interleukin-1 inhibition.
The therapeutic composition of the present invention is ]preferably administered parenterally by ~. .

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injection, although other effectiVe administratlon forms, such as intraarticular înjection, inhalant mists, orally active formulations, or supposit:orias, are also envisioned.
One preferred carrier is physio]Logical saline solution, but it is contemplated that other pharmaceutically acceptable carriers may also be used. In one preferred embodiment it is envisioned ~hat the carrier s3nd the IL-lra constitute a physiologically-compatible, slow-release formulation. The primary solvent in such a carrier may be either aqueous or non-agueous in nature. In addition, the carrier may contain other pharmacologically-acceptable excipients for modifying or maintaining the pH, osmolarity, viscosity, clarity, color, sterility, stability, rate of dissolution, or odor of the formulation. Similarly, the carrier may contain still oth~r pharmacologically-acceptable excipients ~or modifying or maintaining the stabili~y, rate of dissolution, release, or absorp~ion of the IL-lraO Such excipients are those substances usually and customarily employed to ~ormulate dosages for parenteral administration in either unit dose or multi-dose form.
once the therapeutic composition has been ~ormulated, it may b~ stored in sterile vials as a soluti~n, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored either in a ready to U58 form or requiring reconstitution immediately prior to administration. The preferred 5 orage of such ~ormulations is at temperature~ at least as low as 4 C and preferably at 70 C. It is also preferred that such ~ormulations containing Il~lra ar~ stored and administered at or near physiological pH. It is presently believed that storage and administration in a formulation at a high pH
~i.P. greater than 8~ or at a low pH (i.e. less than 5) i5 undesirable.
Preferably, the manner of administering the formulations containing IL-lra is via an intraarticular, ~3~

subcutaneous or intramuscular route. Pr~ferably, the manner of administering the formulations containing IL-lra is via intra-articular, subcutaneous, intramuscular or intravenous injection, suppositories, enema, inhaled aerosol, or oral or topical routes. To achieve and maintain the desired dose of IL-lra, repeated subcutaneous or intramuscular .inj ections may be administered. Both of these methods are intended to create a preselected concentration range of IL-lra in the pa~ient's blood stream. It is believed that the maintenance of circulating concentrations of IL-lra of less than OoOl ng per ml of plasma may not be an effective composition while the prolonged maintenanca of circulating levels in excess of 100 ug per ml may have undesirable side effacts.
A pre~erred dosage range ~or the treatment of interleukin-l mediated arthritis is between 1 and 100 ng/ml.
Accordingly, it is preferred that, initially, doses are administered to bring the circulating levels o~ IL-lra above 10 ng per ml of plasma and that, thereafter, doses are administered at a suitable frequency to keep the circulating level o~ IL-lra at or above approximately lO ng per ml of plasma. The fre~uency o~ dosing will depend on pharmacokinetic parameters sf the IL-lra in the formulation us~d.
A preferred dosage rang for the treatment o~
interleukin-l mediated IBD is between ~bout .5-50 mg pex kg of patient weight administered between about 1 and lO times per day. In a more preferrad embodiment the dosage is between about l-10 mg per kg of patient weight administered between about 3 and 5 times per day. ~he frequency of dosing will depend on ph rmacokinetic parameters of the IL-lra in the formulation us~d.
A preferred dosage ranga for the treatment o~
interleukin-l mediated septic shock i5 between about 1.0-200 mg per kg par day of patient body weight per 24 hours administered in equal doses ~etween about 4-15 times per 24 ~16-2~39~

hours. In a more pre~erred embodiment the dosage is between about 10-120 mg per kg per day of patient body weight administered in equal doses eve:ry 2 hours. In the most preferred embodiment 100 mg per kg o~ patient body weight per 24 hours is equally administered every 2 hours. The frequency of dosing will depend on pharmacokinetic parameters of the Il-lra in the formulation used.
In an additional pref~erred mode ~or the treatment of interleukin-1 mediated septic shock, an initial bolus injection of Il-lra is a~ministered ~ollowed by a continuous infusion o~ IL-lra until circulating IL-1 levels are no longer elevated. The goal of the treatment is to maintain serum IL-lra levels between 2-20 ug per ml ~or this period.
In a preferred emhodi~ent of this mode, an initial bolus of between about 10-20 mg per kg of patient body weight of IL-lra is administered followed by the continuous administration of IL-lra of between about 5-20 ug per kg of patient body weigh~ per minute until circulating IL-l levels are no longer elevated, SBrUm IL-1b level~ may be ascertained by co~mercially available i~munoassay t st kits.
The initiation of treatment ~or IL-l me~iated septic shock should be ~egun, under either mode of treatment, as soon as possible after septicemia or the chance of septicemia is diagnosed. For example, tr~atment ~ay be begun immediately following surgery or an accident or any other event that may carry the risk of inltiating ~eptic shock.
A preerred do~age range for the treatmPnt of interleukin-l m~diated i~shemia and reperfu~ion injury is between abou 1-50 mg per kg of patient weight administered hourly. In a prefarred e~bodiment an initial bolus of about 15-50 mg per kg o~ lra i9 administered, ollowed by hourly injections of about 5-20 mg per kg. The frequency of dosing will depend on pharmacokinetic parameters of the IL-lra in the formulation used.
It is also contemplated that certain formulations 9 ~

containing IL-lra are to be admini~tered orally.
Preferably, IL-lra which is adm.Lnistered in this fashion is encapsulated. The encapsulated IL-lra may be formulated with or without those carriers customarily used in the compounding of solid dosage fo~ns. Preferably, the capsul is dPsigned so that the active portion of the formulation is released at that point in the gastro-intestinal tract when bioavailability i5 maximized and pre-systemic degradation is minimized. Additional excipients may ~e included to facilitate absorption of th~ IT.-lra. Diluents, ~lavorings, low melting point waxes, vegetable oils, lubricants, suspending agents~ tablet disintegrating agents, and binders may also be employ~d.
When used for the treatment of interleukin-l mediated IBD, the administration o~ IL-lra may also be accomplished in a suitably ~ormulated enema.
Regardless of the manner of administration, the specific dose is calculated according to the approximate body weight or surface area of the patient. Further refinement of the calculations necessary to deter~ine the appropriate do~age for treatment involving each of the above mentioned formulations is routinely made by those of ordinary skill in the art and i5 within khe ambit of tasks routinely performed by them without undue experimentation, especially in light of ~he dosage info~mation and assays disclo~ed herein. These dosages may be ascertained through use of the established assays for dete~mining dosages utilized in conjunction with appropriate dose-response data~
It should be noted that the IL-lra formulations described herein may be used for veterinary as well as human applic~tions and that the term "patient" should not be construed in a limiting manner. In the case of veterinary applications, the dosage ranges should be the same as speci~ied abovs.
It is understood that the application of teachings 2 ~ 3 ~

of the present invention to a specific problem or Pnvironment will be within the capabilities of one having ordinary skill in the art in light of the teachings contained herein. Examples of representative uses of the present invention appear in the following examples.
In Examples 1-3, IL-lra's are shown to prevent and/or neutralize, either totally or in part, the ePfects of IL-1 in known arthritis models such as those described by Wilder, R.L., in "Experimental Animal Models of Chronic Arthritis," Goodacre, J.A. and W.C. Dick (ed.), Immunopathogenic Mechanisms of Arthritis, Kluwer Academic Publishers; Dordrecht, Netherlands; Boston, ~ass. (1988), specifically incorporated herein by reference. As is noted in the examples, in each model the IL-lra tested showed beneficial results.
In Examples 4-S~ IL-lra's are shown to prevent and/or neutralize, either totally or in part, the effects of IL-l in known IBD models such as thosD described by Zipser e~ al. Gastroenterology~ 92, pp. 33-39 (1987), specifically incorporated herein by this re~erence; and Sartor e~ al.
Gastroenterology, 89, pp. 587-95 tl985), specifically incorpora~ed herein by ~-his referenr~. As is notPd in the examples, in each model the IL-lra tested showed beneficial results.
In Example 5, IL-lra is shown to have beneficial ef~ects on extraintestin~l inflammation associated with the Sartor IBD model. In Example 6, intestinal inflammation resulting ~rom the administration of an NSAID--indomethacin-~is also neutralized by treatment with IL-lra.
In Example 7, IL-lra's ar~ shown to prevent and/or neutralize, in part, th~ effects of IL-l in a known septic shock model. Rabbits are given intravenous injections QP
endotoxins--believed to be a principal cause of septic shock--to incluce septic shock. The mortality rate of groups ~ 0 3 ~

of rabbits given varying amounts of IL-lra were examined.
As can be seen in the Example, treatment with Il-lra showad beneficial results.
In Example 8, IL-lra's are shown to neutralize, in part, the ef~ects of IL-l in ischemia and reperfusion injury experiments. Dogs are subjected to regional myocardial ischemia for two hours and then reperfused for four hours.
The size of infarct as a perc~ntage of ventricular mass and the in~arct size as a percent of mass at risk were measured.
Treatment with Il-lra showed beneficial result~.
X~MPL~ l: DemonstratiQn of_the E~eçts of Human InteF~eukin~l Inhi~itQx_i~_Cultured Bovine Nasal Cartilaae 2xplant.
Numerous in vitro and in~ivo methods have been used to study the progression of arthritis. One in vitro model which has proven to be especially usaful in this regard is cultured cartilaginous tissue explant. In fact, this model has been used in the past to demonstrate that IL-1 is a powerful mediator of cartilage destruction and, tharefore, a propitiouC target or intervention in arthritic joint erosion. (See qen2rall~ G. Buchan ~t_al., Third Annual General ~eeting of the British Society for ~heumatology, London, England, Novamber 19-21, 1988, PR. J.
Rheu~atol 25 (Supplement 2) 1986; Fontana ~_31-, ~heumatol Int (1982) 2:49-53; J. Saklatvala ~ ., D velopment of Diseases of Cartilage and Bone Matrix, Alan R. Liss, Inc., pp. 291-298; P. Stashenko e~ al., The American Association of ImmunolQgists, Vol. 138, ppO 1464-1468, No. 5, March 1, 19B7; G. Dodge et al., J. Clin. Invest., 83:647-~61; J.
Sandy et al., Journal of Orthopedic Research 4:263-272, J.
Saklatavala et al., The Control of Tissue Damage, Glauert, ed., Elsevier Science Publishers, pp. 97-108; I. Cambell e~
~l., Biochem. J. 237:117-122; J. Tyler, ~iochem. J., 225:493-507, and J. Eastgate et al., Sixth International 2~39Ari~

Lymphokine Workshop, 7(3):338, all of which are specifically incorporated her~in by reference.) The cartilage explant model essentially as described by Steinberg et al., }3iochem. J. 180:403-412, incorporated herein by referPnce, was used in this Example to demonstrate the mitigating e~fect of IL-lra on IL-l mediated cartilage breakdown. ~Yhile bovine nasal septum was used here as the source of cart:ilaginous tissue, articular cartilage of the type described in J. Tyler ~_3~., Br. J.
Theumatol. 24 (Supplement 1):150-155, incorporated herein by re~erence, could also be used.
Preparatio~_~ Cartilaae.
Bovine nasal septum was removed from freshly slaughtered yearling steers and plac~d on ice. The tissue was thPn scrubbed with Povidone/Iodine prep solution (1-ethanol-2~pyrolidinone homopolymer with iodin~, obtained from Medline Industries, Mundelein, Ill). The mucosa and perichondrium were then removed. The remaining cartilaginous septum was then immersed in a 5% (v/v) solution of Povldone/Iodine for one hour at room temperature.
The following procedures were then performed aseptically in a laminar flow hood. The septa were repeatedly rinsed with Gay's Balanced S~lt Solution (GIBC0 Laboratories, Grand Island, NY). The cartilage sheet was then placed on a sterile sur~ace, and uniSorm 8 mm plugs were removed using a standard coxk borer. Approximately 1-2 mm of the top and bottom surfaces were removed using a razor blade. The plugs were then held in the Gey's Balanced Salt Solution~ Plugs taken fro~ different steers were kept separately.
Each plug was then sectioned into several 0.8 mm disks. The cutting devicP used was an aluminum block o~ the type described by Steinberg e~_al.~ Biochem. J. 180:403-412.
The disks produced were consistently be ween 40 and 50 mg wet weight. The dis~s were kept in culture in Delbecco's Modified Eagle Medium plus 10% Fetal ca:l~ serum, plus Penicillin, Streptomycin, and Neomycin (all reagents ~rom GIBCO), hereinafter re~err~d to simply as ~Imedium.~ The cultures were maintained in a 37 C incubator with 5% C02.
Representative disks :Erom each steer were then tested for their ability to respond to IL-lB as indicated by the release of glycosaminoglycans (GAG) into the culture medium. (glycosaminoglycans are released ~rom a cell once ~he cell matrix has been degraded.) The presence o~ GAG was detected using 1, 9-dimethylenethylene blue as described by Ferndale et al., Connective Tissue Research, 9:247-248, incorporated herein by reference. Disks that responded to 5 ng/ml of IL-lB by increasing output of GAG two fold or greater as compared to an unstimulated basal rate were selected for use in th~ following experiments. These disks are hereinafter referred to as "IL-lB responsive disks."
IL-l_Dose Res~nse.
This preliminary experiment was performed to determine whether a dose response curve 0xists to increasing amounts of IL-lB.
First, several of the Il-lB responsive dis~s were sactioned into quarter slices. (The remainder was set aside for later experiments.) Because re~ponses to IL-1 frequently vary from animal to animal, disk to disk, and slice to slice, the steps o~ this e~periment were designed so that each slice serv2d as its own control.
Second, each slice was incubated in one well of a 48 well tissue culture clu~ter tCostar, Cambridge, MA) with a constant volume of the previously described medium. After 48 hours, the amount o~ GAG present in the supernatant of each culture was measured. This amount was then normalized for each culture in terms o~ ug GAG per mg wPt weight of tissue. In this manner, a basal rate of GAG release in the absence of II.-l was established for each slice.

~39~

Third, the supernatants from all the cultures were discarded and replaced with fresh medium containing differing amounts of IL-lB. The IL-lB was produced in-house (J. Childs, notebook 935, pages 49-52) and after characterization, was utilized in all experiments calling for its use. After a 48 hour incubation with IL-lB, the supernatants from the cultures were recovered, and the amount of GAG present in each was measuredO These amounts were normalized for each culture as above. The ~asal rates were then subtracted ~rom the IL-lB inducad rates. The results are depicted in Fig. 1. ~The results are also expressed in tabular form in Table 1.) As Fig. 1 clearly indicates, the r~lease of GAG from th~ cartilaginous tissue is dependent on the amount of IL-lB administered.
Because 5 ng/ml o~ IL-lB caused an easily measurable increase in GAG releas~ during the 48 hour period of culturing, this concentration was used in the following experiment.
Effects of rlL-lra on IL-l Induced GAG Release.
Several of the remainin~ IL-~B responsive disks were n~xt sectioned into quartar slices. As above, each slice was used at its own control.
Each slice wa5 then incubated with a constant volume of the previously-described medium for 48 hours in a 48 well tissue culture cluster. A basal rate of GAG release was determined for each slice. Next, the supernatants ~rom the cultures were discarded and rP.placed with fresh medi~m containing 5 ng/ml o~ IL-lB and difPering amounts of recombinantly produced IL-lra (rIL-lra). A~ter a 48 hour incubation, the supernatants were recovered, and the amounts of GAG were measured. The~e amounts were normalized for each cul~ure by dividing the rIL-lra/IL-lB stimulated GAG
release rate by the basal GAG release rate. The results are depicted in P'ig. 2. As Fig. 2 clearly illus~rates, the release of GAG from cartilaginous tissue was sharply -~3-~ ~ 3 ~

curtailed by an increase in the concentration of rIL-lra relative to that of IL-lB. For instance~, a ten times molar excess of rIL-lra over IL-lB (the molecular weights of IL-lB
and rIL-lra are both approximately 17 kD) was sufficient to return the GAG release rate to the basal level. Similarly, a 1.5 times molar excess of rIL~-lra over Il-lB was sufficient to reduce the stimulation of GAG release to 50%
of that observed in the presence of IL-lB alone.
These results were reproduced using car~ilage derived ~rom several different ~steer~
Lack of Cytotoxiclty of rIL-l~a.
To show that rIL-lra is noncytotoxic, the inventors took slices from the remaining IL-lB responsive disks and ~xposed them to varying amount~ of rIL-lra in the absence of IL-lB. The rate of GAG release was the same as wher~ neither rIL-lra nor IL-lB was pre~ent.
Next, to show that the effects of IL-lra are reversible, ~he inventors then removed rIL lra from the supernatants of culturing slic~s and administered IL-lB
thereto. The slices responded just as they did in thP IL-lB
dose response experiment. Similar results occurred when cartilage that had been treated with IL-lB and a suf~icient concentration of rIL-lra to completely block the action o~
IL-lB was subsequently exposed to IL~l~ alone.
ample 2: Demonst~ation o~ the Effects of Human Inte~leukin 1 Inhlbitor_on Colla~en-Induced Ar~hrit~s in ~i~e.
Type II collagen-induced arthritis in mice bears many resemblances to human rheumatoid arthritis and has been used for several years to study certain aspects of that disease. J. Stuart t al., The FASEB Journal, Vol. 2, No.
14, pp. 2950-2956, Nove~ber 198S, inco~porated herein by reference. The potential involve~ent o~ 1 in rheumatoid arthritis has been noted by S. StimpRon e~ al., The Journal 21~3~5~

of Immunology, Vol. 140, pp. 2964-2969, No. 9, May 1, 198~, also incorporated herein by re~erence.
The purpose of this experiment was to demonstrate that systemic administration of rIL-lra has a mitigating effect on the pathogenesis of type II collagen-induced arthritis in mice.
Twenty-four mice DBA/1 mice, purchased from Jac~son Laboratories, were immunized with 0.1 mg chicX type II collagen in Freund's complet~. adjuvant. At day fourteen post immunization, the animals were randomly subdivided into two groups of twPlve animals each~ The experimental group was injected intraperitoneally twice daily with approximately 0.1 mg rIL-lra/kg/injection~ The injections continued until the animals were sacrificed at day 47 post immunization (i.e., after 34 days of dosing). Control animals were injected with an equal volume o~ vehicle tlO mM
sodium phosphate, 150 mM sodium chloride~ on the same schedule.
Affected limbs were counted and cl.inical scoring was performed approximately three times weekly during the in~ e portion of the experiment. Clinical scores from each animal represent, on a 0-4 point basis, the severity o~
arthritis sustained by each paw as assessed by blinded obserYers. The clinical scores for each animal from day 2SI
when the first sign~ of clinic lly obserYable arthritis were notad, through day 47, when the animals w~re sacri~iced, axe presented in Table 2. The tally of affected limbs and the total clinical score for each group are also pr~sented in Tabla 2. These results are graphed as a function of time in Figs. 3 and 4, respectively. As can clearly be seen, the incidence and severity o~ the disease were slowed down considerably by the administration of rIL-lra.

~3~

Demonstra~ion of the_Effects of Human Interleukin-l Inhibitor on Streptococcal Cell Wall (SCW)-Induc~d Reactivatlon of SCW-Ind~ced Ar~hritis in Rats Regarding streptococcal cell wall-induced arthritis, R. L. Wilder in Immunopathoqenetic Mechanisms o~
a~h~i3i~, Chapter 9 entitled "~Experimental Animal Models of Chronic Arthritis" comments "th~e clinical, histological and radiological features of the ex]perimental joint disease closely resemble those observed in adult and juvenile rheumatoid arthritis".
The experiment described below employs the model disclosed in Esser, et ~1. Arthritis and Rheumatism, 28:
1401-1411, 1985, specifically incorporated herein by reference. This model is briefly summarized as follows:
Streptococcal Cell Wall (SCW~ is injected intraarticularly into the ankle joint of Lewis rats. Saline is injected into the contralateral joint to provide a control. After a period of twenty days, in which the initial inflammation dies away, SCW is again administered, this time by intravenous injection. Thi-~ dose of SCW is insufficient to cause joint inflammation by itself and, therefor~, has little or no ef~ect on the saline injected ankle. In contrast, however, this dose is capable of reactivating inflammation and joint destruction in the ankle previously injected with SCW. To assess the extent of in~lammation following the second administration o~ S~, the dimensions of the ankle jaint are ~ea ured daily.
In one o~ many experiments per~ormed with the above described model, two groups of twelve rats were used.
Each animal was injected in the right ankle with SCW (1.8 ug rhamnose equivalence) and in the left ankle with an equal volume o pyrogen-free saline Ankle dimensions were measured on days 1 through 6.
On day ~0, one group o~ rats was in~ected ~ ~ 3 ~

intraperitoneally with 1 mq/kg IL-lra in an aqueous vehicle;
the other group was injected intrap~ritoneally with an equal volume of the vehicle solution only. One hour later, each animal wa~ injected intravenously with SCW (100 ug rhamnose equivalence). Ten minutes later, the treatment group was injected intraperitoneally with 1 mg/kg Il-lra, and th~
control group was injected with vehicle alone. Subcutaneous injections of Il-lra at 1 mg/kg were given at 2 and 6 hours post-SCW administration and were repPated every 6 hours therea~ter for the next 3 days.
Table 3 and Figure 5 show the dimensions of the saline injected and SCW injected ankles for both the treatment group and the control group over the course of the experiment. As expected, the SCW treated ankles in both groups swelled in responsa to the intravenous in~ection of SCW. However, the re~ponse differed between treatment groups. The ankles in the rontrol group swelled by about 30~ of their initial dim~nsions over the first 3 days whereas the ankles in the treatment group swelled only by 14% over the same period. ~oreover, on days 1 through 5 post-intravenous injection o~ SCW, there was a statistically s~gnificant (P ~ 0.001 by a two-tailed t-test for independent means) difference in thQ dimensions of the SCW-treated and contralateral control ankle~ both groups.
On day 8, the rats were sacrificed and both ankle were fixed in for~alin. The fixed joint~ w~re decalcified, stained, and examined. Significant differences in cartilage erosion, bursiti~, periostitis, and synovitis were found between the control group and the treat~ent group. 50me of these differences are s2t forth in Table 4.
~3~elg_~: Demonst~ation of the E~fects o~ Human In~erleukin-l Inhibitox on Formalin-Immune Com~lex_Induçed IBD.
The rabbit model of formalin~i~mune complex IBD

-~7-~39 ~s~3~

has been used to investigate the role of arachidonic acid-derived inflammatory mediators and to evaluate therapeutic strategies in IBD. Zipser et al. supra; Brown ~_31- 1987 Gastroenterology 92:54-59; Schwnert et_al. 1988 Prostaglandins 36:565 577, incorporatsd herein by reference.
The experiment described below employs the model disclosed in Zipser et al. suPril. This model creates symptoms analogous to active Ulrerative Colitis, and is briefly summarized as follows: formaldehyde is administered via a catheter into the colon of rabbits and after a period o~ time the animals receive an injection of immune complexes in antigen exces~. Time studies following the induction of IBD are conducted by sacrificing the animals after 48 hours and removing the colons. The colons are then histologically assessed. The effect o~ treatment with IL lra's prior to and after the ind~ction of IBD on in~lammation, edema and necrosis was compar~d with non-treated control animals.
nduction of I~D.
Inflammation was induced in the distal colon of male N~w Zealand rabbits ~2.2-2.5kg) using a modification of the immune complex method of colitis Kirsnew et al. 1957 Trans. Assoc~ Am. Physicians 70:lO2-llg; Hodgson et al. 1978 Gut 19:225-32, which are incorporated herein by reference.
Four ml of 0.45% (v/v) unbu~fered fo~maldehyde (Electron Microscopy Sciences, Washington, PA) was administered via a cath~ter inserted 10 cm lnto the distal colon of anesthetized rabbits (xylazine and ketamine). Two hours later, animals r~ceived 0.85 ml o~ immune complexes in antigen excess through an ear vein. The complexes were prepared by incubating human serum albumin (500ug/ml) with rabbit antihuman antisera (ICN Immunobiologicals, Costa Meas, CA), decanting the supernatant, and redissolving the pr~cipated immune complexes with an albumin solution (6mg/ml) as described in Zipser ~ . supra.
Histologic evaluation was per~ormed on a minimum -2~-2~3~

o~ two longitudinal sections from each colon. All colon samples were examined in a blind fashion by a single pathologist. The mucosa and submucosa were separately evaluated for infiltration o~ acute inflammatory cells (neutrophils and eosinophils). A semiquantitive score of leukocytes (L) per high power f:ield (HPF) was determined ~or each area examined usinq the following quantitations: 0 = O
or 1; 0.5 - 2-9; 1 - 10-20; 1.5 = 21-30; 2 - 31-40; 2.5 =
41-50; 3 = 51-65; 3.5 = 66-80; 4 = > 81 L/HPF. At a minimum, eight HPFs of mucosa and submu~osa from each specimen were s~parately evaluated in each section. The inflammatory index was calculated by adding the averaged score for the mucosal and submucosal evaluations. Edema was semiquantitively assessed on a scale o~ 0 to 4. Necrosis was expressed as the percent of mucosa involved. After the administration of formalin, followed by immune complexes, the distal colon develops acute inflammation. This is characterized by in~iltration of n~utrophils primarily into the mucosa and ~ubmucosa, mucus depletion, crypt abscesses, edema and scatt2red areas of mucosal nscrosis, progressively increased from 0.3+0.1 (0 hrs) to 4.5+0.7 ~48 hrs) (p<0.001), from 0.3~0.1 to 3.6+0.3 (p<0.001) and from 0% to 89%(pcO.001) respectively. A subsequent decrease in these parameters was observed 96 hours after the induction of IBD
(p~0.01 versu~ 48 hours).

T~ea~en~ Yith II- lra ' ~ .
A group of animals were treated intravenously with IL-lra (5mg/kg; n=8) or the vehicle alone (n=10) at six time points: 2 hours before and 1, 9, 17, 25, 33 hours a~ter the administration of the immune complexes. The rabbits were sacrificed 48 hours after the induction o IBD and the colon tissue analyzed ~or inflammation.
Treatment o~ rabbits with IL-lra ~ignificantly r2duced inflammatory index from 3.2~0.4 to 1.4~0.3 (p<0.02~, edæma from 2.2+0.4 to 0.6~0.3 (p<0.01) and necrosis from 43~10~ to 6.6~3.2% (p~0.03) compared to vehicle-treated IBD
animals, FIG. 6~ This result shows that several of the indications of IBD may be significantly lassened by treatment with IL-lra.

Exam~le 5: Bacterial_Cell Wall_Induced IBD in Rats Unlike many other IBD models, the bacterial cell wall induced IBD model shows most o~ the indications for chronic IBD or Crohn's disease. In addition to the formation of chronic granulomatous response, this model is subjPct to spontaneous reactivation, anemia and extraintestinal inflammation.
The Bacterial Cell Wall model essentially as described by Sartor ~ . supra., was used in this Example to demonstrate the mitigating affect of IL-lra on IL-l m~diated IBD. The experiment was per~ormed generally as follows: the IBD is induced in rats by the intravenous in;action of a sterile sonicate of peptidoglycan polysaccharide ~rom group A streptococci. Transient petechial hemorrhage o~ the colon appears within 2-3 minutes and resolves by 48-72 hours a~ter injection. A sa~.nplQ group of animals were tseated with IL-lra following induction of I8D, and after a period of time the animals were sacrificed, the colons removed and gross pathology evaluated.
I~duçtion 0~
The ~acterial cell wall material was prepared according to the procedur~s set forth in Stimpson et al.
1986 I~s~ ~Y~- 51:240-249, incorporated her~in by this reerence. -Lewis rats are given subsercosal injections with Streptococcal Cell Walls. The injections result in both local and sy~temic disorders that include bowel adhesions and nodules, an increased liver weight and hepatic nodules, a reduced hematocrit and hemoglobin level, and increased white blood cell count (WBC), a reduced growth rate, and a 4 ~ ~

joint swelling characteristic of arthriti~ (see AppendiX
Sartor, et al., Gastroenterology, 89:587 5g5, 198S, incorporated herein by reference). Three separate protocols for treatment with Il-lra were per~ormed with this model and reductions in nodules and adhesions have been observed in all o~ them; in the last two protocols the reductions in adhesions were statistically signiicant.
Protocol_A.
Two groups of 12 rats were us~d. On day 1, both were inj2cted with 15 ug total of Streptococcal cell wall-derived peptldoglycan polysaccharidP (SCW PG-APS), at seven sit~s; three areas o the cecum, 2 areas of the Peyer's patche~, and two areas o~ the ductal ileum. On ~ay ll overt signs of the disease appeared including joint swelling, diarrhea, and bloody nose. At thi~ time one group was dosed subcutaneously with IL-lra (8 mg/kg3 every 12 hours and the second group was treated identically with placebo (PBS). On each day the size of the ankle joints were measured. On day 18 the animals were sacrificed and the inte~tines were scored on a scale o~ 0 to 4 for the presence of granulomas and adhesions (Table 5). The IL-lra group had fewer nodules and adhesions. The IL lra group also had smaller livers.
Th~ IL-lra group had a reduced whits blood cell count (WBC~.
p~otocol B.
The protocol was similar to that used in protocol A ~xcept th~t the amount of PG-APS used was reduced to 12.5 ug and th~ treatment with lL-lra wa~ started at day 8. A5 in protocol A, reductions in secal nodules, intestinal adhes.ions, liver weights, and WBC were observed (Table 6).
The reduc ion in adhesion was si~nificant at the p ~ 0.02 level.
Proto~ol C.
The protocol used was again similar to that in protocol A except that the amount of PG-APS was reduced to 12.5 ug (as in protocol 3) and the traatment yroup was ~39~

started on IL-lra 8 mg/kg subcutaneous and 2 mg/kg i.v.
i~mediately following the PG-APS injection. Further Il-lra injections t8 mg/kg) s.c. wsre given at 4, 10 and 18 hours on day 1, every 8 houx~ on day 2, and then every 12 hours for the duration of the experiment. FiYe animals in each group were sacrificed at day 3, and th~ remainder were sacrificed at day 18 for examination of gut lesions (Table 7). on day 3 there was a signi~icant reduction in a global parameter representing gut lesions and a reduction in adhesions that approachad significance (p = O.07). In the gxoup sacrificed at day 18 the resultc were confused because no disease appeared in one of the animals in the control group. However, the reduction in adhesions in the IL-lra group was still significant at the p ~ 0.02 level and there was also a signi~icantly greater weight gain in the IL-lra group.

~XAM~LE S: ~SAI~ Induced IBD in Ra~s.
In an attempt to determine whethPr the anti-inflammation effects of IL lra would be additive with those of NSAIDs, rats were treated with indomethacin after the intravenous injection of PG APS as described in Example 3 above (2 n~g/kg/ at the tims o~ reactivation at 12, 24 and 36 hours post activation, and every 12 hours up to 6 days), Il-lra (2 mg/kg at 2 and 6 hours, then eve~y 6 hours up to 36 hours and every 12 hours up to 7 days) or a combination of the two drugs (Figure 7). The group on indomethacin alone showad a greatar reduction in joint swelling than that on IL-lra alone. However, the indomethacin group was sick and two animals died during the course of the experiment. Th~
group receiving both drugs did even better than the group on indomethacin alone; the joint swelling was less, and the difference between the two group was statistically significant on day 4 at p ~ 0.03 and on day 7 and 8 at p~
0.06. No animals were sick in this group and there were ' "~

2 ~ 8 fewer ulcerations in the mid small intestineS. Ulceration of ths mid small intestine is a complication in patients on chronic oral NSAIDs. It appears, therefore, that IL-lra alleviates some of the IBD-like complications of NSAIDs.
Table 8 shows the e~ects o~ IL-lra on both the intestinal symptoms--ulc~rs, adhesions, intestinal t~ickening and myleloperoxidase (MPO) levels--and systemic symptoms -hematocrit (~CT), hemoglobin (HgB) and WBC
levels--assoclated with the N~AID treatment of PG APS
induced arthritis.
xample 7: Demonstration o~ the ~ects of Human Interle~kin-1 ReceRtor Antagonist on Endotoxin Induced S~tic Shock.

Endotoxin induced septic shock studies were conducted on Blue Chinchilla rabbits. The experimental protocol did not ~ocu~ on any indications of the induced septic shock other than group mortality. R~bbits were u~ed in the study because their sensitivity to pyrogenic and metabolic effects of endotoxin and oth~r bacterial products are similar to those of human subjects.
Shock was induced by a single intravenous injection of endotoxin at time zero. ~he rabbit- were giv~n periodic intrav~nous inj~ction~ into an ear ~ein at -10 min., at time zaro, and fvr every two hours therPafter ~or a 24 hour period. The result~ of this study can be seen in TablQ 9.
In Tahle 9, Group A rabbits (n=5) were not given any endotoxin at time zero, and were ~iven saline injections free o~ IL-lra at the periodic injection times. Group B
rabbits ~n=lOj were given .5 mg per kg of ~ody weight of endotoxin at time zero, and the periodic injections were again free of IL-lra. After 7 days the survival rate o~
ra~bits in Group 3 waq only 20%.

2~3~

In Groups C-E (n=10) endotoxin was administered at time zero, and the saline injections contained varying amounts of IL-lra. The rabbits in group C received a total of 10 mg per kg of body weight of IL-lra. The rabbits in group D received a total o~ 30 ~g per kg of body weight o~
Il lra. And finally, the rabbi'ts in group E received a total o~ 100 m~ per kg o~ body weight of IL-lra. After 7 days, the survival rate of rabb.its in group E was 90~.
This experiment, graphically illustrated in Figure 8, shows that treatment with IL-lra signi~icantly delays and reduces final mortality rates in rabbits with endotoxin induced shock.
xam~le 8: ~e~onst~atiQn Q~ t~ g~ct~ of Human Interleukin L Receptor_~nta~onist on Ischemia and Re~r~sion Iniur~
In the following example exp~ximental dogs were subjected to regional myocardial ischemia for two hours and then reper~used for four hours. The doys were divided into two groups, one group treated with IL-lra and the other treatsd with serum albumin i~ the same buffer used ~or the tes~ group.
Animals were fasted ovsrnight and on the ~ollowing morning, were anesthetized with 10 ml o~ thiamylal sodium 5~, followed by 2 ml o~ sodium pentobarbital 6%, intravenously. Additional sodium pentobarbital was admi~istered during the experiment ~s necessary. Arti~icial r~spiratio~ wa~ ~aintained with a Harvard respirator. A
le~t thoracotomy was performed through the fifth intercostal space and polyvinyl catheters placed in the la~t internal jugular vein for fluid and drug administr2tion, and in the lsft internal carotid art~ry and femoral arteries for pressure monitoring and withdrawal of reference blood samples. A c,atheter wa~ placed in the left atrium for in~ection of radio active ~icrospheres. The lsft circumflex artery was dissected free of surrounding tissue and an electromagnetic ~low probe was placed on the vessel proximal to tne ~irst obtuse marginal branch. A~ter an in~ravenous bolus injection of 50 mg of lidocaine, the circumflex coronary artery was occluded w:ith the snar~ occluder for 2 hours. Complete occlusion was verified with the electromagnetic flow probe. The snare was ~hen released suddenly, allowing reperfusion of the coronary vascular bed for 4 hours.
Two-dimensional echocardiogra~s and hemodynamic measurements ~heart rate, blood pressur~ and le~t a~rial pressure) was d~termined before occlusion, after llO min o~
occlusion, 5 minutes after reperfusion, and 4 hours a~ter reperfusion. Two-dimensional echocardiography was performed with the usa of a scanner and a 2.25 MHz transducer. The transducer was placed on the closed shaved right chest and was allowed full visualization of the circumferential extent of the l~ft ventricle in a short~axis pro~ection.
Echocardiographic images were recorded at the midpapillary muscle position onto a video cassatt~ with use of a Sony recorder. A two-dimensional echocardiographic analysis was per. ormed with th~ use o~ A minicomputer-based ~ideo di~itizing system.
End-diastolic and end systolic frames were selected for analysis wtth th~ use of the onsPt of the Q
wave in lead $I as a marXer o~ end-diastole and the smallest le~t ventricular cavity size a~ a marker of end-systole.
Endocardial and epicardial borders for 3 consecutive beats during normal sinus rhythm was care~ully traced directly from the video display onto a digiti2ing tablet. Quantitive analysis was performed with a radial contraction model and a fixed diastolic center of mas~ at 22.5 d~gree intervals over the full left ventricular circumference.
The midpoint of the posterior papillary muscle was chosen as a :Eixed anatomic re~erence and designated as 135 2 ~ 3 .~

degree~. Wall thickening was computed ~or each of the 22.5 degree sectors with the ~ollowing e~uation: wall thickening = [(end systolic wall thickness - end diastolic wall thickness)/ end diastolic wall thickness~ x 100%. The normal range of wall thickening was determined from a functional map of the baseline images for three cardiac cycles and 95% tolerance limits were establi hed in each animal. These limits were used ~or comparison with occlusion and reperfusion functi.onal maps and abnormalities are expressed as the circumfererltial extent of dys~unction and the degree of dysfunction. The ~xtent of dysfunction (in degrees~ was measured at th~ intercepts between the occlusion or reperfusion maps and ths low4r 95% tolerance limit; the degree of dys~unctio~ (in area units) is the planimetered area below the lower ~5~ tolerance limit.
Regional myocardial blosd flow was assessed by the reference withdrawal ~ethod using tracer-tabled microspheres (15 ~m diamater, New England Nuclear) injected into the left atrium. The microsphere~ were ultrasonicated and vortex-agitated before injection. ~icrospheres were injected before occlusion, a~ter 110 ~in of occlusion, 5 minutes after reperfusion and 4 hours aftar r perfusion with one of six available isotopes ll4lCe, 51Cr, 113Sn, 103Ru, 9sNb, 46Sc)~
Simultaneous re~erenc~ arterial samples were withdrawn fro~
the carotid and femoral arteries at ~ constant rate of 7 ml~min with à Harvard withdrawal pump starting 10 sec before microsphere injection and continuing for 120 sec after completion o~ th~ injection.
Two adjacent transverse left ventricular slices at the midpapillary muscle level, corresponding to the echocardiographic short-axis slices, were selected for blood flow determination. Each slice was divided into 16 full thickness 22.5 degree sectors. ~ach sector was then ~urther divided into epicardial, midmyocardial, and endocardial samples. The tissua samples were then weighed, placed in counting vials, and assayed for radioactivity in a gamma scintillation count~r. A~ter background and overlap correction, absolute myocardial blood ~low was calculated with the following equation: Qm=(Cm x Qr/Cr), where Qm =
myocardial blood ~low (ml/min); Cm = counts/ min in tissue sample; Qr = withdrawal rate of the re~erence arterial sample (ml/min); Cr - counts/min in the reference arterial sampl~. Myocardial blood ~low i5 exprQssed per gram of tissue ~or each sample.
Just prior to sacri~ice, the le~t circumflex coronary artery was briefly occluded and monastral blue pigment (0.5 ml/kg) injected into the left atrium for d~lineation of the in vivo myocardial area at risk. The animal th~n received 3000 U o~ heparin and was sacrificed with an intravenous bol~s of saturated KCI solution and the heart excised.
~ reatmen~ Gr~u~s. Dog were randomly assigned to one o~ two groups. In the test group, dogs received a bolus injection oP ~0 mg Il-lra inhibitor just prior to the onset of the ischemia and 15 mg IL-lra inhibitor for each hour until the expariment was terminated. Control animals ~eceived an identical ~u~ntity of endotoxin-~ree, human albumin dissolved in the same buffer used ~or the test group.
~ 5~ Y5I~3~5~ A. A~ter death, the heart oP each dog was 2xcisQd,the l~ft ventricle i~olated rom surrounding tissue, cooled in a ~reezer for 15 minut~s, and then sliced into 5 mm transverse sections. The slices were then weighed and placed in a warm bath of bu~ered triphenyl tetrazolium chloride ror ten minutes. In th s t~chnique, viable tissue stains red whil~ nonviable tissue remains unstained (A~. Heart. J. 101:593). The unstained zone of infarcted ti~ue i6 outlined on transparent overlay~
and quantitat:ed by planimetry u~ing ~ microcomputer and corrected for the weight of the heart slice. Infarct size is expressed as the percentage of the area of myocardium at risk (the area at risk of infarction is de~ined as the area of the myocardium left unstained following the injection o~
monastral blue into th~ left atrium).
NMR Analysis of myca~dial_~d$~. After fixation, the hearts were cut into 5 to 7 transverse slices approximately 5 mm thick. Two transmural myocardial tissu~
samples were obtained ~rom the nonischemic zone (positive msna~tral blue staining) and the central ischemic zone (n~gative blue staining). The ~picardium for each sample was dissected away to eliminate possible lipid signal interf~rence. Each piece was subdivided transmurally (weighing approximately 500 mg ~ach) with one portion assessed for % H20 by desiccation te~hnique (wet weight -dry weight/wet weigh~ whil~ the other wa~ placed into a clean dry glasc tubeO T1 and Tl relaxation times were obtained on a IBM PC 20 Minispec spe~tro~et~r (IBM
Instruments, Inc., Danbury, CT) operating at 20 ~Hz and 40C. The location of the sample in the magnet, 90 and 180 radio frequency pulses, and detector phase were o~timized for each sample before relaxation measurement~
w~re ob~ained. ~1 valu~s w~r~ determir~ed by a ~it o~ 20 inversion data recovery points while T2 values were d~termined by using a Carr, Purcell, Meiboon-Gill (CPMG~
sequence. In an attempt tQ minimiza effects of di~usion and mi~cellan~ou~ system instabiliti~, the 180 radio frequency interpulse spacing was maintained at 180 microseconds. The fraction of echo samples determined were used as variables to ad~u~t the duration of the CPMG
experiment. Typically, 1 to 150 data points were acquired as the ec:ho train was delayed to ~ 5% to 25% of its original amplitllde. T2 values were determined by using a multi exponantial i~it. Only the dominant component of the 2xponential ~Eit was used ~or 3tatistical analysis. The r~sults o~ the Tl and T2 analysis were corrected with . ..

2~3~

percent water for adjacent tissue samples to veri~y the accuracy o~ the NMR technique.
~ _loqic and m r~hometr.ic ~valuat1ons. For each group tested at least 3 animals were ~valuated by light microscopy. Sections stained with hematoxylin and eosin from each heart were evaluated ~or n~utrophil accumulation within the area betwe~n viable and in~arcted tissue.
Stat stic AnalYsis. ~11 data was represented as -thP mean -~ SEM. Comparisons within groups were made by a two-way analysis o~ variance; when significant F values are obtained, paired t tests (corrected ~or multiple comparisons with the Bonferroni inequality adjustment) will be us~d ~o determine which maasurem~nt dif~ered significantly ~rom one another.
Comparison~ between groups were made by unpaired t test. An exponPntial regression was used to correlate infarct size data to myocardial blood flow.
The results of the I~-lra treatment regimen on protecting dog myocardium from occlusion reperfusion injury, are listed in Table 10 below. As a percentage o~ the left ventricular ~as~ the in~lux infarct size in the treated group was reduced to 10.3% as opposed to 18.~ in the control animals. This represen~s a 40% reduction in the percent of tha left ventricular ~as~ that was in~arcted.
The percentage of the area at risk in contrast~ was not markedly chan~ed, 40.5% of t~e left ventricular mass in th~
treated ~roup versus 44.8% in th~ control animals. When the in~arcted area is calculated as a perc~nt o~ the total area at ri~k, the numbers similarly-favor th~ IL-lra treated animals, 24.3~ versus 42% in the control group.

-39~

' ' .

:`

The effect of IL-lra in reducing the extent at infarcted tissue in canine coronary occlusion-reperfusion studies.
ILlra Treated ~ 91 ~I~LY~ LL~L}~L~9 Infarct si~e as a % of le~t ventricular mass 10.3%-+2.2% 18.2%-~3.3%
Area at risk as a % of left ventricular mas~
40.5%-+1.7% 44.8%-+1.9%
Infarct size as a of mass at risk 24.9%-+4.6% 42%-~.3%
Al~hough the present invention has been de~cribed in connection with preferred embodiments, it is understood that those skilled in ths art are capable of making modificatisns and variations without departing from the scope or spirit of the present invention. Therefore, the ~oregoing d~scription of prefPrred e~bodim~nts is not to be taken in a limiting sense, and the present invention is best d~fined by the ~ollowing clai~s and their equivalents.

-4~

-4~ 2 0 3 9 ~ 5 8 The ef~ect of recombinan~ IL~ on ~L-1,B induce~ degradation of Bovine l~ds~l ~, ~n all cases ~IL 1~] -; ng/ml.

(~L-1QJ [IL-LGl Fold Stirnulation ~IL-lb] penod lI/periodl (~/- standard deviation) 0 4.02 ~/- 1.7 n~/rnl 1 2.4 ~/- 0-47 10 ng/rnl 2 1.7 ~- 0.4 2~ ng/ml 5 1.3 +/- 0.4 50 n~/ml 10 1.0 +/~ 0.2 1~0 ng/ml 30 1.1 +/ 0 2 . .

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-~l3 -Table 3 ANKLE JOINT DIAP~IETER OF RATS INJECTED
WITH SCW AND TREATED WIT11 IL~1rz.0R SALINF
ACCORDING TO PROTOC:OL IN EXAMPLE 3 Joint Diameter (mm) (~_) SCW Injected Joints Saline In~ected Joints__ . SQ ~aline ~ Sl:) Saline SD
0 5.96 .1 2 6.02 .10 5.95 .17 5.96 .1 6 7.95 .33 7.73 .36 5.94 .15 5.94 .13 2 7.44 .28 7.42 .27 5.98 . l 1 5.95 .17 3 7.~0 .39 7.23 .27 6.00 .1 2 6.01 .07 6 6.78 .27 6.64 .29 6.06 .09 6.06 .13 6.5~ .34 6.63 .18 6.00 .12 5.85 .16 14 6.44 .21 6.36 .17 5.99 .08 5.90 .17 6.46 .18 6.52 .14 5.91 .11 5.87 .~0 21 7.34 .36 7.78 .31 5.73 .18 5.78 .12 2? 8.31 .58 8.70 .43 5.85 .16 ~.96 .22 23 8.55 .B1 9.06 .42 6.02 .19 5.99 .16 24 8.23 .71 8.56 .39 6.03 .13 5.94 .20 8.00 .56 8.16 .43 6.05 .12 6.06 .17 28 7.48 .4~ 7.71 .30 ~.04 .13 5.98 .13 ~,~3~

Table ~ -4'1-EHects ot IL-1rGon Joint Histopathology Following SCW P~activation of Joint Inflammation (1 rng/kg 4 times daily on day 20 through 23) Placebo Group IL~ Group P
.
PalhologyPositives/ 12 ScorePositives/ 12 Score .
Cartilage Erosion 10 1.0 + .6 3 0.25 + .45 .0023 8Ona Erosion 3 0.25 ~ .45 2 0.17 + .39 NS

Bursitis 11 0.92 ~ .29 3 0.25 ~ .45 .0003 Periostitis 9 0.75 +.4512 0.25 + .45 .013 Synovitis 12 2.21 ~ .84 12 t.08 ~ .47 .OOû52 PMN t2 1.0 12 1.0 NS
.

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Tabl e Effects of IL-lra on Scw ~ nduced enterocolitis in the rat Intestinal Cecal Liver W~C
Adhesions ~Q~ ~Qi.ghS_~
IL-lxa 1.7 1.816.9 48.8 P~S 2.2 2.41a.6 57.7 p Yalua 0. 14 0 . 1n o. lg o .13 Por comparison o~ grs--ps -- r 2~3~ 4~ ,3 Table ~ -q6-Effects of IL-lra on SCW-induced enterocolitis in the rat Intestinal Cecal Liver W~C
~dh~sions ~Q~lÇ~ Weiqht ~ImL
IL-lra 1.4 1.7 13.3 35.1 P~s 2.2 2.3 14.1 35.~
p value 0.017 0.077 0.23 0.43 rOr co~pariaon o~ groups : - .

~, . .
: ::
.

. . .

~39~

TablQ :;a -~7-Effects of IL-lra on SCW-induced enterocolitis in the rat .
Intestinal Cecal Liver WBC
Adhçsio~s ~Q~ isht (qm~ -IL-lra 0.8 0.9 0.047 10.7 P~S 1.8 1.0 0,049 10.3 p valua 0.07 0.30 0.24 0.31 for comparlson of group~
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~ xperlmental en~o~oxln In~uc~ ~hoc~ in rab~lts; etfects ~ IL-1 ra on surYival rate sur~ival (no) ~survival rate 7 days (%) 12h 24h 36h 48h 7d A (N-S) ~ 5 5 5 5 1~0 8 (N~10) 9 6 3 2 2 ~0 C (N~10) 9 7 4 3 2 20 1: (N=1~) 10 7 6 5 . 40 (N=10) 10 10 10 9 9 90 ;

Claims (32)

1. A medicine for treating interleukin-l mediated disease of a patient in need thereof, which comprises a therapeutically effective amount of an interleukin-l inhibitor in admixture with a pharmaceutically acceptable carrier.

2. The medicine of claim 1 wherein the interleukin-l inhibitor is a protein.

3. The medicine of claim 1 wherein the interleukin-l inhibitor is IL-lra.

4. The medicine of claim 3 wherein the IL-lra comprises at least one compound from the group consisting of IL-lraa, IL-lrab, IL-lrax and methionyl Il-lra.

5. The medicine of claim 4 wherein the IL-lra is IL-lraa.

6. The medicine of claim 4 wherein the IL-lra is IL-lrab.

7. The medicine of claim 4 wherein the IL-lra is IL-lrax.

8. The medicine of claim 4 wherein the IL-lra is methionyl IL-lra.

9. The medicine of claim 4 wherein the IL-lra is produced by a recombinant DNA method.

10. The medicine of claim 9 wherein the IL-lra is produced in substantially pure form.

11. The medicine of claim 1 which is in a liquid form.

12. The medicine of claim 1 wherein the interleukin-l inhibitor is an IL-l binding protein

13. The medicine of claim 12 wherein the IL-l binding protein is a soluble receptor.

14. The medicine of claim 12 wherein the IL-l binding protein is a monoclonal antibody.

15. The medicine of claim 1 wherein the interleukin-l inhibitor blocks IL-l production.

16. The medicine of claim 16 wherein the interleukin-l inhibitor that blocks IL-l production is IL-lra.

17. The medicine of claim 1 wherein the interleukin-1 mediated disease is selected from the group consisting of:
arthritis, inflammatory bowel disease, septic shock, ischemia injury, reperfusion injury, osteoporosis, asthma, insulin diabetes, myelogenous and other leukemias, psoriasis and cachexia/anorexia.

18. A medicine for preventing interleukin-l mediated disease of a patient in need thereof, which comprises a therapeu-tically effective amount of an interleukin-l inhibitor in admix-ture with a pharmaceutically acceptable carrier.

19. The medicine of claim 18 wherein the interleukin-1 inhibitor is IL-lra.

20. The medicine of claim 19 wherein the IL-lra is selec-ted from the group consisting of IL-lraa, IL-lrab, and IL-lrax.

21. The medicine of claim 18 wherein the interleukin-1 inhibitor is an IL-l binding protein.

22. The medicine of claim 21 wherein the IL-l binding pro-tein is a soluble receptor.

23. The medicine of claim 21 wherein the IL-l binding protein is a monoclonal antibody.

24. The medicine of claim 18 wherein the interleukin-1 inhibitor blocks IL-l production.

25. The medicine of claim 24 wherein the interleukin-1 inhibitor that blocks IL-l production is IL-lra.

26. The medicine of claim 18 wherein the interleukin-1 mediated disease is selected from the group consisting of:
arthritis, inflammatory bowel disease, septic shock, ischemia injury, reperfusion injury, osteoporosis, asthma, insulin diabetes, myelogenous and other leukemias, psoriasis, and cachexia/anorexia.

27. A medicine for preventing or treating interleukin-1 mediated disease of a patient in need thereof, which comprises a therapeutically effective amount of an interleukin-1 inhibitor in admixture with a pharmaceutically acceptable carrier.

28. A medicine of any one of claims 1 to 27 which is in a dosage unit form containing the interleukin-1 inhibitor in such an amount that from 0.01 ng to 100 µg of the interleukin-l inhibitor is contained per ml of plasmain patient blood stream when administered.

29. A medicine of claim 28, wherein the amount is from 10 to 100 ng per ml of plasma and the disease is interleukin-l mediated arthritis.

30. A medicine of claim 28, wherein the amount is 0.5 to 50 mg per kg of patient weight; the medicine is adapted to be administered 1 to 10 times per day; and the disease is interleu-kin-l mediated IBD.

31. A medicine of claim 28, wherein the amount is 10 to 120 mg per kg per day of patient body weight; and the disease is interleukin-l mediated septic shock.

32. A medicine of claim 28, which is in a commercial package that carries indications that the medicine can be used for preventing or treating interleukin-l mediated disease.