MXPA00004583A - Treatment of kaposi's sarcoma with il-12 - Google Patents

Abstract

Methods are provided for using IL-12 to treat Kaposi's sarcoma (KS), particularly AIDS-associated KS.

Description

* TREATMENT OF KAPOSI SARCOMA WITH IL-12 Field of the Invention The present invention relates generally to the treatment of Kaposi's sarcoma 5 ("KS"), particularly KS associated with AIDS, with interleukin-12 ("IL-12").

Background of the Invention This application is a continuation in part of provisional application serial number 60 / 100,416, filed September 15, 1998, which is incorporated herein by reference. reference. Reports in 1981 of Kaposi's sarcoma (KS) associated with AIDS in homosexual males increased the interest in this until now rare neoplastic disease. It soon became clear that the KS associated with this epidemic was more fulminant than any of the classic or African or endemic forms, generally following a more progressive course. quickly. At the onset of EDIS, KS may progress to involve viceral organs and, particularly pulmonary KS, it is not infrequent to cause death. Furthermore, aggressive cutaneous and lymphatic involvement is frequently an important cause of morbidity. At one point, KS was the second most common defining disease of AIDS. However, as the epidemic has matured and the definition of what AIDS constitutes has been modified, the incidence of KS as the initial defining disease of AIDS in patients has collapsed. However, the absolute number of cases of KS continues to increase and KS now often develops after patients have another AIDS defining disease. The distribution of KS in the establishment of AIDS is not uniform among all groups at risk for HIV infection, with most of the cases occurring in male white homosexuals. In fact, KS is seven times more common in homosexual or bisexual men (27.3%) than in all other AIDS patients combined (3.9%). There is currently no curative therapy for KS. While there is some evidence to suggest that antiretroviral therapy may, under certain circumstances, delay or even reverse Partially the development of KS, this tumor usually requires a specific therapy.

Many modalities have been tried with different results. Interferon alfa has been useful in obtaining good responses, particularly in patients with disease limited to the skin and with T4 cell counts that are above 200 / mm3. However, this is not a cure, and interferons can have toxicities and often overlap with those of AZT and can interfere with antiretroviral therapy. Currently, ongoing studies try to administer interferons with several antiretroviral therapies. Localized KS lesions are usually treated with radiation therapy. More aggressive or viceral lesions are usually treated with cytotoxic chemotherapy. Chemotherapy of KS with various agents has resulted in tumor responses that may be important; however, these responses tend to be incomplete, temporary and often shorten life. Also, many of these agents are associated with significant immunosuppression and myelosuppression, and patients often can not tolerate therapy for long periods of time. In addition, treatment with many of these agents may predispose patients to the development of opportunistic infections (Oi). Although the pathogenesis of KS is not completely understood, there is an important body of evidence that shows that the process of angiogenesis is central to the initiation and spread of KS lesions. Cells derived from Kaposi's sarcoma and cultured in vitro have been shown to secrete a variety of autocrine and paracrine growth factors, including some with potent angiogenesis including basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and interleukin-1 (IL-1). Many of these same factors, in addition to others such as the dissemination factor (SF) and HTV-1 Tat, have also been shown to stimulate the in vitro growth of spindle cells derived from KS. These same KS cells, when inoculated subcutaneously in nude mice, were found to be capable of inducing the growth of a lesion similar to KS that was originated from mouse tissues. In addition, similar lesions were induced by the co-inoculation of bFGF and Tat as well as Tat and heparin. In this way it would appear that the spindle cells derived from KS secrete factors that, alone or in combination with Tat, are capable of inducing the formation of lesions similar to KS. In addition, the growth and development of these lesions in mice can be inhibited by the systemic administration of agents or compounds with angiogenic activity such as the tissue inhibitor of metalloproteinase-2 (TIMP-2) or tecogalan. There are also data implicating endothelial cells as the cells of origin for spindle cells of KS. Cultured endothelial cells acquire a fusiform morphology when exposed to cytokines released in vitro by activated T lymphocytes, including many of the cytokines that induce in vitro proliferation of spindle cells derived from KS. In addition, these endothelial cells also become sensitive to the in vitro proliferative effects of Tat in a manner similar to that of spindle cells derived from KS. The initial stimulus of the factor that initiates the proposed cytokine cascade leading to the development of KS is not known. However, there is a growing body of literature suggesting that a virus similar to herpes, tentatively called herpes virus Kaposi's sarcoma (KSHV) or human herpes virus 8 (HHV-8) may be involved. There are reports that this virus appears to be present in or associated with KS lesions from HIV infected as well as from classical and African KS with a much higher frequency than in KS-free patients, although it has been reported in a number of other diseases such as lymphomas in body cavities and Castleman's disease. In addition, there is a recent report that KSHV is present in the vascular spaces of the flat endothelial cell lining of KS lesions as well as in spindle cells typical of KS. Thus, although the exact mechanism and stages associated with the development of KS are not known, it is clear that angiogenesis is central to the overall pathogenesis of this disease. Studies have shown that CD4-positive T lymphocytes can be divided into two main groups: T-type 1 (THI) helper cells, which produce interleukin-2 (IL-2) and interferon-gamma.; and helper cells T type 2 (TH2), which mainly produce interleukin-4 (JL-4), interleukin-5 (IL-5), interleukin-6 (IL-6) and interleukin-10 (IL-10) 1"4. In general, THI cells mediate type-1 cellular immune responses, whereas TH2 cells are involved in humoral type-2 immune responses. In the establishment of HIV infection, there is a decrease in IL-2 production and other reactions mediated by THI type 1 with an improvement in TH2-mediated reactions, type 2, and this change in immune responses away from the predominance of type 1 is thought to be the central feature of the HIV infection.5'6 In summary, the KS associated with EDS is a serious neoplastic disorder that is associated with AIDS, which can cause significant morbidity and even death.The treatment of this disorder is currently not optimal, and there is a definite need for new agents that are better As noted above, there is evidence that neovascularization is important in the development of KS. For this reason, it is hypothesized that the inhibition of angiogenesis may potentially benefit patients with KS. As will be described below, interleukin-12 (IL-12) has been shown to have potent anti-angiogenic activity and can substantially stimulate THI immune responses. Interleukin-12 (IL-12) is a cytokine that has been found to enhance the proliferation of natural killer (NK) and activated T cells; improves the cytotoxic activity of T and NK cells; and induces the production of interferon-gamma (IFN -?) 7. Also, IL-12 has demonstrated anti-tumor and anti-metastatic activity in preclinical models. In addition, IL-12 has been shown to potentiate the growth and differentiation of THI cells while inhibiting the activity of TH29. This effect of the differentiation of the T-helper through IL-12 has also been found to restore the immune responses mediated by HIV-specific cells ex vivo in T cells from patients infected with HIV10. More recently, Voest and colleagues have reported that EL-12 is a potent antiangiogenic agent11. They also found that the anti-angiogenic activity of IL-12 was mediated through the induction of IFG-α. Subsequently, Angiolillo and colleagues reported that the human protein capable of being induced by interferon (IP-10), a chemokine induced by IFN-α, is a potent inhibitor of angiogenesis in vivo12. Therefore, it seems that the anti-angiogenic activity of EL-12 can be through its induction of IFN-α production. local with subsequent overregulation of IP-10 production. Thus, IL-12 has been shown to possess potent anti-angiogenic activity in vivo as well as selective improvement of THI activity. Foli and associates have recently reported the effects of IL-12 on HIV replication in vivo13. They demonstrated that E-12 induced in vivo replication of HIV in fresh and pre-stimulated peripheral blood mononuclear cells (PBMC) as determined by the production of HIV p24 antigen in 7 days of culture. This replication of HIV induced by IL-12 was not attributable to the induction of IL-1, IL-2, tumor necrosis factor alpha or beta and was associated with the selective loss of the CD4 subgroup in stimulated cultures. However, EL-12 had little or no effect on the replication of HIV in monocytes / macrophages. Finally, they demonstrated that the increase in HIV replication induced by IL-12 could be inhibited by the dideoxy-nucleosides AZT, ddl and ddC. Thus, there is the possibility that the systemic administration of IL-12 may increase the replication of HIV in infected patients, particularly in the absence of antiretroviral therapy. There have been numerous Phase I and Phase II trials administering IL-12 in intravenous or subcutaneous form and using a variety of doses and schedules. A variety of toxicities were noted. These included symptoms of fever and constitutional; nausea, vomiting anorexia, diarrhea, stomatitis, dyspepsia and positive guiacal stools; transient increases in leukocyte, lymphocyte, neutrophil and platelet counts with anemia and occasional increases in PT and PTT; transient increases in serum glucose values and less frequent hyperglycemia; dose-dependent elevations of SGOT, SGPT, alkaline phosphatase and bilirubin; dyspnea, hematuria, proteinuria, elevated serum creatinine and elevated blood urea nitrogen, and oliguria; confusion, anxiety, fading, insomnia, hypotonia, nervousness, drowsiness, and tremor; hypotension and some peripheral edema; erythema and pruritis at the site of subcutaneous injections that subside spontaneously; and increases in thyroid stimulating hormone (TSH). Six patients died while receiving EL-12; 4 deaths were from the progression of the disease, while 2 were attributable to IL-12. Two patients whose death was thought to be related to IL-12, both were enrolled in Phase II trials that treated patients with renal cell carcinoma with 500 ng / kg of IL-12 intravenously daily for 5 days every three weeks. One patient died on day 10 of the cycle as a result of a low gastrointestinal hemorrhage, and the other died on day 23 of cycle 1 due to multiple organ failure. Finally, it was determined that the toxicity profile of IL-12 was highly dependent on the program and that the dose administered in the Phase II trial was too high for that program. In particular, the first Phase I studies had used a single dose of EL-12 with a delay before multiple doses were given, whereas the Phase II trial did not use this first single dose. There are data that suggest that through an as yet undetermined mechanism, this initial dose of IL-12 induced a type of tolerance that reduced the toxicity of subsequent doses. There is information on Phase I, single-dose, placebo-controlled studies in which EL-12 is administered subcutaneously to patients infected with HIV, either with 100-300 CD4 cells / mm3 or 301-500 cells CD4 / mm3 . The dose levels studied were 3, 10, 30, 100, 300 and 1000 ng / kg. It was found that the maximum tolerated dose (MTD) was 300 ng / kg based on the observed toxicity at 1000 ng / kg (fatigue, stomatitis, elevations of aminotransferase, and hyperbilirubinemia). There were no significant clinical changes in CD4 or CD8 cell counts or plasma viraemia (as measured by a branched-chain DNA assay) in any subject. There were, however, transient increases in the levels of interferon-gamma and neopterin observed in patients who received doses greater than 100 ng / kg. A Phase I / II, double-blind, randomized, placebo-controlled, multicenter, and increasing-dose trial of IL-12 in HIV-infected patients who had 100 to 500 CD4 / mm3 cells was started in November 1995. It is administered interleukin-12 by subcutaneous injection twice a week for 12 weeks. The dose levels to be studied are 30, 100, 300 and 500 ng / kg / injection of IL-12. Twenty-four patients will be enrolled in each cohort (18 receiving IL-12 and 6 receiving placebo) to ensure that there will be 20 patients susceptible to evaluation who will end at least 4 weeks of study drug administration. A total of 26 patients have been enrolled at the dose level of 30 ng / kg. Four patients have been withdrawn from the study for reasons related to non-safety. Thirty patients have completed 12 weeks of therapy. The dose of 30 ng / kg has been tolerated well. To date, no serious adverse events have been reported that are related to the administration of the study drug or dose-limiting toxicities. The escalation of the dose up to the dose level of 100 ng / kg has occurred and 10 patients have been enrolled at this dose level.

It would be desirable, therefore, to provide treatment for KS by administering IL-12. OBJECTIVES OF THE INVENTION The present invention provides a method for the treatment of Kaposi's sarcoma in a mammalian subject, which method comprises administering to the subject a therapeutically effective amount of EL-12 or a subunit or biologically active fragment thereof.

In other embodiments, the present invention provides a method for inhibiting angiogenesis in a lesion associated with Kaposi's sarcoma in a mammalian subject, which method comprises administering to the subject a therapeutically effective amount of IL-12 or a biologically subunit or fragment. active of her. In preferred embodiments, the treated subject has AIDS. In other preferred embodiments, EL-12 is administered as a protein or in the form of a DNA encoding IL-12. In still other preferred embodiments, the EL-12 protein is administered in a dose of from 100 to 1000 ng / kg body weight of the subject, preferably in a dose selected from the group consisting of 300, 500, 625 and 750 ng / kg body weight of the subject, and more preferably at a dose of 300 ng / kg body weight of the subject. Other aspects and advantages of the present invention will be clear from the consideration of the following detailed description of the preferred embodiments thereof.

Detailed Description of the Invention Interleukin-12 (EL-12) originally termed the natural killer cell stimulating factor, is a heterodimeric cytokine described, for example, in M. Kobayashi et al., 1989, J. Exp. Med. 170 : 827 EL-12 can be purified from natural sources, produced by chemical synthesis, or preferably produced by recombinant DNA techniques, for example, by the expression and isolation of EL-12 protein in recombinant host cells as described in detail in the international patent application WO90 / 05147, published on May 17, 1990 (also European patent application No. 441,900), incorporated herein by reference. The DNA and amino acid sequences of the 30 kD and 40 kD subunits of heterodimeric human IL-12 are provided in the aforementioned international application and in the US patent. No. 5,571,515, incorporated herein by reference. Research quantities of recombinant human and murine IL-12 are also available from Genetics Institute, Inc., Cambridge, Massachusetts. As used herein, the terms "interleukin-12" and "IL-12" refer to interleukin-12, its individual subunits and fragments thereof that exhibit adjuvant activity of EL-12, polynucleotides encoding EL-12 , and functional equivalents of "interleukin-12" and "IL-12." A therapeutically effective amount of IL-12 is an amount that when administered results in (1) decreased symptoms of KS, including, without limitation, a decrease in the number, recurrence, spread or size of one or more KS lesions; or (2) a reduction in angiogenesis in one or more existing KS lesions or in the formation of new KS lesions. The amount of IL-12 administered to the host will vary depending on a variety of other factors including the antigen (s) used, size, age, body weight, general health, sex and diet of the host, the time or duration of the administration and the particular qualities of the KS lesions that are being treated. As an example, a therapeutically effective amount of IL-12 polypeptide is desirable between about 10 ng to about 1000 ng (preferably about 100 ng to about 750 ng, or about 100 ng to about 300 ng, or about 300 ng to about 500 ng, or about 500 ng to about 500 ng to about 750 ng) of EL-12 polypeptide per kilogram of patient's body weight. The preferred doses are 100 ng / kg, 300 ng / kg, 500 ng / kg, 625 ng / kg and 750 ng / kg. The therapeutically effective amount for any particular patient will be easily defined by balancing the efficacy and toxicity of EL-12 administration. The adjustment and manipulation of established dose ranges are common tasks within the knowledge of those skilled in the art. EL-12 can be administered to a host in a variety of ways. Routes of administration include, without limitation, intradermal, transdermal (e.g., by slow-release polymers), intramuscular, intraperitonal, intravenous, subcutaneous, oral, aural, epidural, anal or vaginal routes (e.g., by suppositories) and the intranasal.

Any other convenient administration route can be used, for example, bolus infusion or injection, or absorption through epithelial or mucocutaneous coatings. Particularly preferred routes of administration are those that patients can conveniently administer to themselves in home settings, such as, without limitation, the intradermal, transdermal (e.g., by slow release polymers), intravenous, subcutaneous, oral routes. , aural, epidural, anal or vaginal (for example, by suppositories) or intranasal, or by absorption through epithelial or mucocutaneous coatings. In addition, IL-12 can be administered in combination with other components or biologically active agents, such as any of a variety of antiviral agents (e.g., nucleoside reverse transcriptase inhibitors, non-nucleoside analog reverse transcriptase inhibitors, and / or protease inhibitors) or any of a variety of antineoplastic or chemotherapeutic agents (eg, acyclic nucleoside phosphonates, Adriamycin, bleomycin and / or vincristine, either individually or in combination), or other biologically active agents such as tretinoin . Other pharmaceutically acceptable components can also be administered in combination with IL-12, for example surfactants such as glycerides, excipients such as lactose, carrier diluents and carriers. If desired, certain sweeteners, flavors and / or coloring agents can also be added. In addition, IL-12 can be administered by in vivo host expression of the nucleotides encoding EL-12. The polynucleotides, preferably in the DNA form, can be released to the patient for in vivo expression of IL-12. The so-called "naked DNA" can be used to express EL-12 in vivo in a host. (Cohen, J., 1993, Science 259: 1691-1692, Fynan, E. et al., 1993, PNAS USA 90: 11478-11482, and Wolff, JA et al., 1991, Biotechniques 11: 474-485 describe similar uses of 'naked DNA', the entirety incorporated herein by reference). Yet other modes of delivering L-12 to the patient in the form of polynucleotides encoding it are known to those skilled in the art and may be employed rather than administration of the IL-12 polypeptides, as desired. For example, polynucleotides encoding IL-12 can be administered as part of a vector or as a cartridge containing the IL-12 coding sequences operatively linked to a promoter sequence, (e.g., see PCT international patent application. WO / 01139, published on January 20, 1994 and incorporated herein by reference). Briefly, the DNA encoding IL-12 protein or desired fragments thereof can be inserted into a nucleic acid cartridge. This cartridge can be manipulated to contain, in addition to the sequence of the EL-12 to be expressed, other optional lateral sequences that make possible its insertion within the vector. This cartridge can then be inserted into an appropriate vector downstream of a promoter, a mRNA leader sequence, an initiation site and other regulatory sequences capable of directing the replication and expression of that sequence in vivo. Additional regulatory sequences may also be inserted downstream of the coding sequence to be expressed. This vector allows the in vivo expression of the IL-12 polypeptides within the host. The patent and literature references cited herein are incorporated by reference as if they had been fully established. The following examples illustrate embodiments of the present invention, but it is not intended to limit the scope of the description.

Example 1 - Clinical Treatment of KS Associated with AIDS, with IL-12 As of the time of this presentation, the following clinical protocol has been and is being carried out in human subjects at the National Cancer Institute. Despite the use of present or future tense in the text that follows, the procedures reported have been and are currently carried out.

Eligibility and Enrollment Evaluation Patients are evaluated for their eligibility according to the following criteria for inclusion or exclusion. Inclusion Criteria: • Patients with HIV infection and Kaposi's sarcoma (KS) confirmed by biopsy who have a disease that is amenable to evaluation by non-invasive methods and a life expectancy of > 3 months.

All patients must also have serum antibodies to HIV as determined by ELISA and western blot. Patients should be ambulatory with a Karnofsky performance status of at least 70. All patients should be receiving a stable dose of antiretroviral therapy consisting of a combination of two or more of the following: AZT, ddl, ddC, 3TC, d4T, saquinavir , ritonavir, indinavir, a non-nucleoside reverse transcriptase inhibitor, or other protease inhibitor. Treatment with other therapies is permissible as long as it is considered a standard of community care. Patients have to be in this regimen for 4 weeks before therapy with IL-12. Patients must be at least 18 years of age and be able to give informed consent. All patients with gestational potential or who have children must agree to use medically accepted means of birth control while they are in the study and for 2 months after this. A total bilirubin of < 3.7 mg / dl with direct fraction of < 0.2 mg / dl and indirect fraction of < 3.5 mg / dl in patients for whom these abnormalities are believed to be due to protease inhibitor therapy.

Exclusion Criteria • Pregnancy, or possibility of pregnancy during drug administration. All female patients with gestational potential must pass a negative pregnancy test within 2 weeks of entry into the study. All patients with gestational potential or who have children should agree to use medically accepted means of birth control while in the study and for 2 months after the study. In addition, patients who breastfeed or breastfeed will not be allowed in the study. In this regard, it should be noted that the greatest risk in breastfeeding is the transmission of HIV to the progeny, with an additional risk due to the study drug.

Patients with pulmonary KS or who seriously threaten life and who can respond to other therapies will be ineligible. In addition, patients with KS lesions with active or critically localized bleeding will be ineligible if, in the opinion of the Principal Investigator or the Study Chair, these injuries pose an immediate risk to the patient. Any of the following hematologic abnormalities: 1. An Hb < 9.0 gm / dl or transfusion within one month before entry. 2. An absolute neutrophil count (ANC) < 750 cells / mm3. 3. A platelet count < 75,000 cells / mm3. 4. APTT or PT > 120% control A history of liver cirrhosis or current hepatic dysfunction with: 1. Total bilirubin > 2 mg / dL. 2. AST / GOT > 2.5 times the upper limit of normal. Serum creatinine > 1.5 mg / dL and an estimated or measured creatinine clearance < 60 mL / min. Clinically important autoimmune disease (ie, disease caused by the production of antibodies and / or an immune response of the body against itself, such as systemic lupus erythematosis), and / or rheumatologic disease. Dense and active gastrointestinal bleeding or peptic ulcer disease without control.

A history of inflammatory bowel disease. Past or present history of malignancies other than KS, except for patients who have been in complete remission during > 1 year after the interruption of their therapies, or that they have completely removed basal cell carcinoma or squamous cell carcinoma in situ of the cervix. Evidence of a severe or life-threatening infection with bacterial, viral, fungal, protozoan, or parasitic pathogens within 2 weeks of entry into the study. In general, patients who have had a > 39 ° C or more within 10 days prior to entry into the study will be ineligible, unless it is evident that this is not due to a severe underlying infection.

Patients with some other abnormality, except lymphopenia or direct manifestations of KS, that would be registered as grade 3 toxicity (see Table 1 below). Known hypersensitivity to EL-12 or other compounds that are known to cross-react with IL-12. Patients who must not have previously received EL-12 for any reason. Also treatment within the last 6 weeks with suramin; treatment within the last 3 weeks with any agent or cytotoxic chemotherapeutic regimen (6 weeks for mitomycin C or nitrosoureas), interferon, other therapy or systemic anti-KS regimen, radiation therapy, or local treatment (such as intralesional injections), treatment within the last 6 weeks with cytokines or bone marrow stimulating factors other than erythropoietin (Epo); treatment within the last 2 weeks with systemic glucocorticoid steroids in sufficient doses to affect the immune response. In general, this would mean an equivalent of more than 20 mg of prednisone for more than 1 week. Replacement glucocorticoid therapy may be allowed. It is not recommended although androgynous or mineralocorticoid therapy is allowed. I? impossibility of giving informed consent or reluctance to abstain from unprotected sexual contact or other activities that could result in reinfection with HIV. Any medical condition that, in the opinion of the Principal Investigator or the Study Chair, would prevent the inclusion of a patient in this research study.

Evaluation of Research Eligibility The pretreatment evaluation will be carried out within 14 days of the initiation therapy and will include a complete history, review of systems and physical examinations, with particular attention to the neurological examination and any neurological malignancy. If the patient is to begin treatment within 48 hours of the selection, the following studies need not be repeated at the time of enrollment. Selection studies will include: • HIV serology. • Severe care panel Hepatic panel Mineral panel CPK, uric acid, amylase, lipase, LDH CBC, diff (automated lymphocyte count), retic count, ESR. APTT, PT, thrombin time (TT), fibrinogen. Urinalysis Pregnancy test (if female) EKG Chest x-rays CD4, CD8, RA and T RO cells including FACS (Four green 10 cc tubes for SIAC, Frederick, MD, scheduled in advance, which must be collected up to before 12 noon, a simultaneous CBC and an automated differential should be obtained). Red tip tube for Frederick, MD for storage. Yellow tip tube for PCT storage of plasma HIV-1 RNA (send to SIAC, Frederick MD).

Patient Registration The participation of each patient will be discussed with the Principal Investigator (Dr. Yarchoan) or with the President of the Study (Dr. Pluda). Once it has been determined that a patient meets the requirements, informed consent will be obtained by the patient, as documented by a signed statement of informed consent, approved by the NCI-ERB. The physician in charge of patient care must register the patient with the Medicine Branch protocol office by calling Orkand Co. In addition, Orkand Co. must be called to the same number at the time a certain patient is removed from the protocol. Prior to administration of the first dose, a history and a general medical examination, as well as a negative pregnancy test, where appropriate, will be recorded, together with an evaluation of the patient's KS.

Maximum number of patients: 55 Implementation of the Study This study is designed as a pilot trial administering 3 successively high doses of IL-12 to patients with KS associated with HIV. Information related to the tolerability of each dose will be collected, as well as preliminary information regarding the activity of EL-12 when administered to such patients. Data will also be collected regarding the immunological and virological effects of EL-12 when administered to patients with KS associated with HIV. Patients will receive IL-12 twice a week subcutaneously (SC). Successive groups will receive successively higher doses of EL-12; 100 ng / kg / dose, 300 ng / kg / dose, 500 ng / kg / dose, 625 ng / kg / dose and 750 ng / kg / dose. However, unless a dose is modified in response to toxicity, a given patient will receive a constant dose of IL-12 without intrapatient dose escalation. Patients can receive IL-12 initially for up to 12 weeks. Previously, patients who achieve a stable or better clinical KS response after 12 weeks could receive up to an additional 16 weeks of therapy for a total of 28 weeks of treatment. Since beneficial antitumor or immunological effects have been observed and these continue after 28 weeks of therapy, an amendment has been sought to extend the period of drug administration to 18 months (year and a half). Additional patients can be treated at the highest tolerated dose in order to explore the activity of EL-12 in patients with KS associated with HIV. Initially, 3 to 6 patients will be entered at each dose level. Up to 10 additional patients can be enrolled at the highest tolerable dose to confirm the tolerability of this novel dose and obtain exploratory information on the activity of IL-12 in this patient population. Up to 3 of the 6 original patients at each dose level who do not complete the initial 4 weeks of therapy for reasons other than drug toxicity can be replaced (for a maximum total accumulation of 55 patients). Patients should not have presence of pulmonary KS lesions, critically localized or with active bleeding. The dose level will be scaled in successive groups of 3 patients as long as dose-limiting toxicity is not observed (see the section on "Toxicity Criteria" below).

If instance 1 of dose-limiting toxicity is observed among the 3 initial patients at a dose level, an additional group of 3 patients should be treated at that dose level without additional dose limiting toxicity in order that such escalation of dose can proceed. If 2 instances of dose-limiting toxicity are observed at a dose level, the highest tolerated dose has been exceeded. It is desirable that at least 6 patients be treated at the previous level to ensure their tolerability. The highest tolerated dose will be defined as the highest dose level administered as part of the protocol where 0 of 6 or 1 of 6 patients (eg, <1/6 patients) experience dose-limiting toxicity (see section on "Toxicity Criteria" later for the definition). In determining the highest tolerated dose level, up to an additional group of 10 patients can be enrolled at this dose level in order to better define the tolerability and toxicity of this dose level and to preliminarily explore the activity of the IL- 12 in patients with KS associated with HIV. It is possible that, after a review of data from this or other studies with administration of EL-12, the accumulation of additional patients at a dose level below the highest tolerated dose can be guaranteed. If this occurs, an amendment to the protocol will be made to allow additional patients to be enrolled at dose levels below the highest tolerated dose.

Drug Administration Patient evaluation and administration of the drug will be performed at the outpatient facilities. However, admission may be required for the evaluation of complications if medically necessary. Patients will receive their initial dose of IL-12 at the Oupatient Cancer Center (NCI Outpatient Cancer Center) (day hospital) and will be observed for 1 hour immediately after administration. Subsequent doses of EL-12 will be administered at the NCI Oupatient Cancer Center, the doctor's office at the patient's home, or at home by a nurse or other professional trained in health care. A single dose of IL-12 will be administered subcutaneously (SC) twice a week, with at least 3 days of separation, to 3 successive groups of patients. The doses of IL-12 administered will be: Level 1 100 ng / kg / dose twice a week. Level 2 300 ng / kg / dose twice a week. Level 3 500 ng / kg / dose twice a week. Level 4 625 ng / kg / dose twice a week. Level 5 750 ng / kg / dose twice a week.

Three to six patients will be initially introduced at each dose level. EL-12 will be administered to patients within each dose level for up to 12 weeks. Up to 3 patients at each dose level who are withdrawn from the study before completing 4 weeks of therapy for reasons other than drug toxicity may be replaced, at the discretion of the Principal Investigator or the Protocol Director. After at least 3 patients have completed the first 4 weeks of administration of EL-12 without developing a dose-limiting toxicity (see the "Toxicity Criteria" section presented below for definition) at a level of At a particular dose, subsequent patients can be enrolled at the next higher dose level. In determining the highest tolerated dose level, up to about 10 additional patients can be enrolled at this dose level in order to better define the tolerability and toxicity of this dose level and to preliminarily explore the activity of IL-12 in patients with KS associated with HIV. Initially, patients who achieve a clinical response to stable KS or better after 12 weeks could receive up to an additional 16 weeks of therapy. Since beneficial antitumor or immunological effects have been observed and these continue after 28 weeks of therapy, an amendment has been sought to extend the period of administration of the drug to 18 months (one year and a half). It has been attempted for at least 6 patients receiving the highest tolerable dose, to have their IL-12 injections applied immediately to one side of a cutaneous KS lesion other than the marker lesion (see section on "Screening").

Kaposi's sarcoma "later for the definition of a marker lesion) for a minimum of 4 weeks.

Modifications to Treatment With the exception of more below elevations of transaminase grade 4 (see Table 1 below), patients experiencing dose-limiting toxicity (see the "Toxicity Criteria" section below for definition) may be re-treated at the next lower dose level (50% of the starting dose for patients in level 1), provided the toxicity is resolved to < grade 1 within 4 weeks of stopping the drug. Patients whose toxicity is not resolved within 4 weeks of IL-12 arrest or who have the recurrence of dose limiting toxicity at a lower level will be withdrawn from the study. Patients who develop dose-limiting transaminase elevations lower than grade 4 and who resolve as indicated above can be treated at the same dose of EL-12. If dose limiting liver toxicity occurs again and is recovered without the drug as mentioned above, then they will be re-treated with the next lower dose. Patients who develop hepatic transaminase toxicity dose-limiting at this lower dose will be withdrawn from the study.

Patients who develop hair or excessively bleeding stools will have their IL-12 retained and pending appropriate evaluations.

Study Evaluation: The following baseline studies are performed before starting treatment (if the patient initiates the treatment within 48 hours after the selection, the evaluations of the list will not need to be repeated in "Evaluation of Eligibility of Investigation" ): Severe care panel Liver panel Mineral panel Thyroid function test (TSH, TT4) CPK, uric acid, amylase, lipase, LDH, beta-2-microglobulin CBC, diff (automated lymphocyte count), retic count, ESR. APTT, PT, thrombin time (TT), fibrinogen. Urinalysis • Pregnancy test (if female) • CD4, CD8, RA and T RO cells including FACS (Four 10 cc green tip tubes for SIAC, Frederick, MD, scheduled in advance, which must be collected before 12 noon, a simultaneous CBC and an automated differential should be obtained). • EBV antibodies, CMV antibodies • Hepatitis B S Ag, Hepatitis B S Ab • Hepatitis C antibodies • RBC folate, vitamin B 12, iron, transferrin, VDRL • Red tip tube for Frederick, MD for storage. • 40 cc of heparinized blood (in green tipped tubes) will be removed for immunological testing (TH1 / TH2 profile and production of IL-12 in response to mitogens and antigens) (send to Dr. Gene Shearer's lab, cubicle 5A31) . • Up to 20 cc can be extracted for virological testing, establishment of cell lines, evaluation of hematological parameters, or other studies that become clinically important during the conduction of the test (If researchers are interested in studying genetic / family markers, they will return to the IRB with an amendment to the protocol in order to carry out this investigation). • Quantitative levels of immunoglobulin, including IgE. • Yellow tip tube for PCR storage of plasma HIV-1 RNA (send to SIAC, Frederick, MD). • Two red tip tubes for interferon-serum range, IP-10, bFGF, VEGF, and IL-6. Place one of these tubes on ice immediately upon extraction (send to Dr. Yarchoan's laboratory, cubicle 5A25). • 20 cc of urine for bFGF (send to Dr. Yarchoan's laboratory, cubicle 5A25). • EKG • CXR; Chest CT whenever possible (CT is required when the clinical scenario or CXR results suggest a pulmonary process). • Other studies, where indicated, to evaluate and measure internal tumors.

It was required to test the KS by biopsy before the patient entered the study. (Patients had to bring slides from an external biopsy for review or have a biopsy done during the selection process). further, a biopsy of an easily accessible skin lesion, normal skin, or samples of pleura or ascitic fluid can be obtained for research purposes, such as the establishment of cell lines and attempts to identify possible viruses related to KS. Refusal to allow such sampling will not prevent a patient from entering the study.

Evaluation of Kaposi's sarcoma Full-body, reference photographs were obtained upon entering the study. At this time, 5 lesions (hereinafter referred to as marker lesions) representative of the patient's disease were chosen and, if possible, located in separate areas of the body. These marker injuries should be lesions that have never been treated with local therapies such as radiation therapy or intralesional injections. Detailed photographs of this lesion will be obtained with a metric ruler on the side of them. The size, color and nodularity of these lesions would be recorded at each clinic visit. If there are a total of more than 50 lesions, 1 to 3 representative areas of the body containing more than 20 KS lesions will be chosen (see also the "Response Criteria" section below). An attempt will be made to distribute the "marker" lesions between the representative areas and the rest of the body. Radiological studies will be performed at the entrance where it is clinically indicated. The evaluation of all the corporal injuries of the KS will be carried out as follows: The patients will be evaluated at the entrance to determine if they have 50 or more injuries. (1) For patients with 50 or more lesions at the entrance, 1 to 3 representative areas will be selected in reference and these will be used for each subsequent evaluation.

Representative areas are sections of the body (for example, back, leg, arm, etc.) that contain at least 20 KS injuries. The total number of injuries in these representative areas will be counted and recorded if they are flat or raised. Yes, in the course of treatment, a single injury is divided into 2 or more smaller lesions (whose area does not extend beyond the initial injury limit), these lesions will still be counted as single lesions for purposes of the evaluation of the total numbers when defining a response to therapy. (2) For patients with less than 50 injuries upon entering, the total number of injuries will be counted and recorded if they are flat or raised.

Phase of Treatment v Monitoring General evaluation. Patients will be evaluated in clinic once a week during the first six weeks of therapy, and subsequently every 2 weeks while in the study. The evaluation will be made at each clinic visit of the subjective symptoms, including headache, nausea, vomiting, diarrhea, abdominal discomfort, appetite, tremor, night sweats, rash, muscle and joint pains, ability to concentrate, paresthesias and status mental and neurological Objective signs will include a complete physical examination, weight changes, and fever. Temperatures and weights will be evaluated at each clinic visit.

Evaluation of Kaposi's sarcoma Evaluation at each clinic visit: Marker lesions will be measured and their size, color and nodularity will be recorded at each clinic visit. Evaluation every 4 weeks: (1) Marker lesions will be measured and a record will be made of their size, color and nodularity, and the total number of lesions (if the patient, at the reference point, subjects less than 50 lesions) or lesions within representative areas previously defined (if the patient had more than 50 lesions) will be counted at the end of every 4 weeks of treatment as described in the "Kaposi Sarcoma Evaluation" section, as well as at the end of treatment or when the patient leaves the study. (2) Full-body photographs and marking injuries will be obtained at the end of every 8 weeks of treatment, as well as at the end of treatment or when a patient leaves the study. Radiological studies, where appropriate, will be repeated every 8 weeks (or more frequently if clinically indicated) as well as at the end of treatment or when a patient leaves the study.

For patients receiving their EL-12 injections immediately adjacent to a KS lesion, this lesion will be photographed, measured and their nodularity and color recorded before initiating therapy. This lesion will be measured and its nodularity and color will be recorded at each clinic visit while the injections continue to be applied adjacent to it, and photographs will be taken every 8 weeks or whenever the injections adjacent to it are discontinued, regardless of whether the patient continues to receive EL-12 injected elsewhere.

Laboratory studies. The following laboratory studies should be obtained at each clinic visit (but not more than once a week, unless clinically indicated): • CBC with differential, ESR reticulocyte count. • APTT, PT, fibrinogen. • Critical panel, liver panel, mineral panel. • Urinalysis.

Tests obtained every 4 weeks: • CBC with differential, ESR, reticulocyte count • APTT, PT, fibrinogen. • Critical panel, liver panel, mineral panel. • Thyroid function test (TSH, TT4). • Red tip tube for Frederick, MD for storage. • Urinalysis, CPK, amylase, LDH, beta-2-microglobulin, uric acid. • Two red tipped tubes for serum bFGF, VEGF, interferon-gamma, IP-10, and IL-6. Place one of these tubes on ice immediately upon extraction (send to cubicle 5A25). cc of urine for bFGF (send to cubicle 5A25). CD4, CD8, RA and T RO cells including FACS (Four 10 cc green tip tubes for SIAC, Frederick, MD, scheduled in advance, which must be collected before 12 noon.) A simultaneous CBC should be obtained and an automated differential). Up to 20 cc of blood can be optionally extracted for virological testing, establishment of cell lines, evaluation of hematological parameters, or other studies that become clinically important during the conduction of the test (Yes researchers are interested in studying genetic / family markers, they will return to the ERB with an amendment to the protocol in order to carry out this investigation). Yellow tip tube for PCR storage of plasma HIV-1 RNA (send to SIAC, Frederick, MD).

Tests obtained at weeks 4, 8, 12 and 24: • 40 cc of heparinized blood (in green tip tubes) will be extracted for immunological testing (TH1 / H2 profile and production of EL-12 in response to mitogens and antigens) (send to Dr. Gene Shearer's laboratory, cubicle 5A31).

Tests at the end of the study: All blood tests required every 4 weeks.

A chest x-ray will be done every 6 weeks while you are in therapy.

Patients can have biopsies of cutaneous lesions easily accessible and obtained as clinically indicated. In addition, at the entrance to the protocol or at any time during the course of the study, a biopsy of a representative lesion of skin not involved in pleural or ascites fluid may be obtained for research purposes, such as the establishment of cell lines and attempts to identify them. of possible viruses related to KS. Additionally, cutaneous lesions not related to KS can be obtained for the same studies. If biopsies of visceral KS lesions are made for clinical purposes, samples may also be used for these research purposes. Biopsies of appropriate KS lesions can be obtained to determine whether a representative lesion has been pathologically resolved. The rejection of the submission of such biopsies will not prevent a patient from entering or continuing in the study. Other tests may be obtained as clinically indicated. The tests can be re-scheduled to the closest possible day without this constituting a protocol violation (for example, Federal holidays, or unforeseen circumstances such as travel problems).

Concurrent Therapies See the section on "Supportive Care" below for a description of antiretroviral therapy. Every effort should be made to give only medications that are clearly indicated for a specific medical purpose during the test. It is particularly important to avoid if systemic administration of glucocorticoid steroids is entirely possible, since its use can exacerbate the KS. In addition, drugs that are likely to affect KS lesions, immunological parameters, or HHV-8 / KSHV should be avoided wherever possible. Any other treatments, medications, biological products, or blood products, including counter medications, imported drugs, or street drugs that the patient has taken in the first month before beginning therapy will be recorded. Any available information related to exposure to drugs or nephrotoxic agents, toxic to the bone marrow, immunomodulatory or hepatotoxic will be recorded. With the exception of the exclusionary drugs mentioned above, patients will be kept generally with the drugs that they have been taking before their entry, unless medically a change in the drug regimen is required. Patients will not receive any immunomodulatory agent or, as noted above, therapies for their KS. Patients will only receive erythropoietin (Epo) or, if absolutely required, a granulocyte colony stimulation factor (G-CSF) according to standard medical practice. The use of other cytokines will not be allowed. Finally, careful attention should be paid to the potential for drug interactions between protease inhibitors and other drugs that the patient may be taking.

Criteria for Withdrawal from the Study Treatment will be discontinued for any of the following reasons: • The occurrence of dose-limiting toxicity. However, patients who experience dose-limiting toxicity (see section "Toxicity Criteria" for definition) can be re-treated with the lowest dose level nearby (50% of the starting dose for patients in the level 1) as long as the toxicity is resolved to the reference point or grade 1 (whichever represents the most abnormal value) within 2 weeks of stopping the drug. Patients whose toxicity is not resolved within 2 weeks after the arrest of EL-12 or who have the recurrence of dose-limiting toxicity at a lower dose will be withdrawn from the study • Patients who develop a neutrophil count absolute (ANC) < 500 cells / mm3 can continue receiving the EL-12 and G-CSF, 300 μg three times a week (tiw), added until the ANC is > 750 cells / mm3. If the ANC does not increase to > 750 cells / mm while receiving the G-CSF, then the EL-12 will be stopped and the G-CSF continued. The EL-12 can then be reset to the next lowest dose level (50% of the starting dose for patients in level 1) if the ANC goes up to > 750 cells / mm3 within 2 weeks of stopping the drug. Patients whose ANC does not increase to > 750 cells / mm3 within 2 weeks of the arrest of EL-12 or those who have recurrence of dose-limiting toxicity at a lower dose level will be withdrawn from the study. An attempt will be made to stagger and discontinue the G-CSF while the ANC stay > 750 cells / mm3. • The appearance of a life-threatening infection after the patient has been enrolled in the study. In this case, the patient can resume the reception of IL-12, at the discretion of the Principal Investigator or the Study Chair, after terminating the therapy for the infection, provided that this period is not longer than 4 weeks. However, those patients who have previously shown evidence of improvement with EL-12 and who have had EL-12 withdrawn for more than 4 weeks may, at the discretion of the Principal Investigator or the Study Director, be placed again with EL-12 within 2 weeks of the resolution of the disease.

• Generalized debilitation of mental incapacity that would render the patient incapable of giving informed consent. • Any other potential adverse reason or event that is considered sufficiently serious by the Principal Investigator or the Study Director to decree the suspension of the therapy. • Therapy may be discontinued due to patient's failure to comply with the protocol, at the discretion of the Principal Investigator or the Study Director. • Pregnancy of a patient. • Inability to take a combination of 2 or more antiretroviral agents as established in the "Supportive Care" section below. • Progress of the disease or disease that is sufficiently severe so that, in the opinion of the Principal Investigator or the Director of the Study, cytotoxic therapy is required. Interleukin-12 can, as with other angiogenesis inhibitors, be cytostatic and therefore result in either stable disease or a decrease in the rate of KS advancement. Therefore, patients with a disease that, although progressive by definition in the "Response Criteria" section below, is not severe or life threatening, may, after discussion with the patient and the Principal Investigator or the Study Director, continue to receive EL-12. • Therapy may be discontinued at the patient's request or at the discretion of the Principal Investigator or the Study Director.

Post-Study Evaluation Patients will be observed for a follow-up evaluation 4 weeks after the end of treatment. If there is any toxicity of the drug in the follow-up evaluation, patients will be followed as medically indicated until the toxicity stabilizes. The evaluation of subjective symptoms, including headache, nausea, vomiting, diarrhea, abdominal discomfort, appetite, tremors, night sweats, rash, muscle and joint pains, concentration capacity, paresthesia and mental and neurological status. Objective signs will include a complete physical examination, weight changes, and fever. Temperatures and weights will be evaluated at each clinic visit.

Laboratory studies: CBC with differential, ESR, reticulocyte count APTT, PT, fibrinogen. Critical panel, liver panel, mineral panel. Thyroid function test (TSH, TT4). Red tip tube for Frederick, for storage. Urinalysis, CPK, amylase, LDH, beta-2-microglobulin, uric acid. Two red tipped tubes for interferon-gamma, IP-10, serum of bFGF, VEGF, and IL-6. Place one of these tubes on ice immediately upon extraction (send to cubicle 5A25). • 20 cc of urine for bFGF (send to cubicle 5A25). • CD4, CD8, RA and T RO cells including FACS (Four 10 cc green tip tubes for SIAC, Frederick, MD, scheduled in advance, which must be collected before 12 noon.) A simultaneous CBC must be obtained and an automated differential). • Up to 20 cc of blood can be extracted optionally for virological testing, establishment of cell lines, evaluation of hematological parameters, or other studies that become clinically important during the conduction of the test. • Yellow tip tube for PCR storage of plasma HIV-1 RNA (send to SIAC, Frederick, MD). • 40 cc of heparinized blood (in green tipped tubes) can be extracted for immunological testing (TH1 / TH2 profile and production of EL-12 in response to mitogens and antigens) (send to Dr. Gene Shearer's laboratory, cubicle 5A31 ).

Supportive Care Medications may be administered as clinically indicated, or at the discretion of the Principal Investigator or the Study Director, with the following exceptions. • Specific therapy for KS during the first 12 weeks of study. After this period, lesions with occasional or disfiguring pain may, in rare cases, be treated with localized therapy at the discretion of the Principal Investigator or the Study Director and the concurrence of the 1ND holder. However, the patient must continue to have at least 5 KS lesions susceptible to evaluation. However, every effort will be made to avoid such therapy. Any medications noted in the exclusion criteria with the exception of short courses of corticosteroids.

All patients should be receiving a stable dose of antiretroviral therapy consisting of a combination of two or more of the following AZT, ddI, ddC, 3TC, d4T, saquinavir, ritonavir, indinavir, a non-nucleoside reverse transcriptase inhibitor, or other inhibitor. of protease, either alone or in combination, for 4 weeks before therapy with IL-12 and while in the study. Treatment with other therapies is allowed as long as it is considered a standard of community care. In general, every effort should be made not to change the patient's antiretroviral therapy during the protocol, while the effect (if any) of IL-12 on viral load is of interest. However, changes in dosage may be made if they are medically required. Also, if medically indicated, a patient must switch between antiretroviral agents or regimens after entering the study, while continuing to receive a combination of 2 or more agents. Patients who require the interruption of their antiretroviral therapy for any reason will be removed from the study. All patients should receive a therapeutic multivitamin with minerals, and low levels of vitamin B12 or folate should be corrected with specific vitamin replacement therapy. Patients should also be in prophylactic therapy against opportunistic infections, as medically indicated.

Data Collection and Evaluation The data will be collected by the members of the Retroviral Disease Research Team. Dr. James M. Pluda, President of the Protocol, will be immediately responsible for omissions of the protocol.

Response Criteria: The evaluation of the KS response to an agent or regimen is difficult to assess by commonly used oncological definitions. Nevertheless, in an effort to standardize the evaluation of KS therapy, the Oncology Committee of the AIDS Clinical Trial Group Oncology Committee has devised a group of stage and response definitions for the KS. We will use a modification of these criteria to evaluate the responses of patients with KS to the administration of IL-12. It should be noted that there is some variability of the observer in the evaluation of the number, size, nodularity and color of the lesions, and this should be taken into account when the measurements are interpreted. For the evaluation of less than complete responses in patients with more than 50 lesions upon entering, only representative areas 1-3 previously selected and containing at least 20 lesions will be considered. However, complete responses still require the absence of any detectable disease on the whole body (that is, not confined to representative areas).

Complete Response (CR): The absence of any detectable residual disease, including tumor-associated edema, that persists for at least 4 weeks. In patients in whom pigmented macular skin lesions persist after apparent CR, biopsy of at least one representative lesion is required to document the absence of malignant cells. In patients known to have visceral disease, an attempt to re-staging should be made with appropriate endoscopic or radiographic procedures. If such procedures are medically contraindicated, the patient may be classified as having a clinical RC.

Partial Response (PR): Sih net increase in the number of injuries (noting, as described in the section "Evaluation of Kaposi's Sarcoma," the simple lesions that are divided into two or more smaller lesions during the course of the treatment will still be counted as one); no new injuries occur in areas of the body previously not involved; without new visceral sites of involvement or the appearance of worsening of edema associated with tumor or effusions; and (1) a 50% or more decrease in the number and / or size of previously existing injuries and lasting for at least 4 weeks or (2) complete flattening of at least 50% of all previous elevated injuries (ie, 50% of all previous nodular plaque type lesions become macular) lasting for at least 4 weeks or (3) A 50% decrease in the sum of the products of the largest perpendicular diameters of the marker lesions lasting for at least 4 weeks or (4) A 50% decrease in visceral lesions that can be measured radiologically as confirmed by a radiologist at the Clinical Center (usually Dr. Fuerstein) sustained without evidence of regrowth for at least 4 weeks or (5) Patients who otherwise meet the criteria for CR but who still have edema associated with residual tumor or effusions, will be classified as having PR.

Complete Clinical Response: The absence of any detectable residual disease, including edema associated with tumor, persisting for at least 4 weeks. For patients with macular and pigmented skin lesions that persist after apparent complete response, a representative lesion has not been biopsied and found to be disease free. For patients with visceral disease, the radiological diagnosis or the endoscopic study should be repeated if it is not medically contraindicated and is found to be negative for evidence of disease.

Progressive Disease: For those criteria that involve the measurement of lesions in the clinic, the designation of progression should be made, when feasible, only in the case that the following criteria have been satisfied with a separation of 1 week. (1) An increase of 25% or more over the reference point in the number of injuries and / or size (sum of the products of the largest perpendicular diameters) of the marker injuries, or (2) A change in the character, from macular to plaque-like or nodular macular of at least 25% of the lesions, or (3) new visceral sites of involvement or progression of visceral disease, or (4) The development of new or growing edema associated with tumor or effusion that lasts for at least 1 week and interferes with the normal activities of the patient.

Toxicity Criteria The NCI Common Toxicity Criteria (CTC) will be used to assess toxicity (see Table 1 below). However, since lymphopenia is almost a universal finding in late HIV infection, lymphocyte counts will be recorded according to CTC but will not count as a dose-limiting toxicity of IL-12 or as a criterion to discontinue therapy. Thrombocytopenia grade 4; grade 4 anemia not responsive to erythropoietin; and grade 4 neuropenia not responsive to G-CSF (see section "Study Withdrawal Criteria" above) will be considered dose limiting. Lymphopenia is a common manifestation of HIV infection and will not be used as a dose-limiting toxicity, no matter what the degree. Neurological and cardiac grade 2 toxicities will be considered as dose limiting. Any grade 3 or non-haematological toxicity (excluding those mentioned in the "Toxicity Criteria" section, hyperbilirubinemia, and elevated liver transaminase levels) will be considered dose-limiting. For patients with increased bilirubin, a total bilirubin > 4.8 nm / mL if direct bilirubin is > 0.3 mg / mL and indirect bilirubin is > 4.5 mg / mL, will be considered as dose limiting. For patients with elevated hepatic transaminase levels, a transminase > 500 IU will be considered as limiting the dose. Any grade 3 or non-haematological toxicity (excluding those mentioned in the "Toxicity Criteria" section and hyperbilirubinemia) will be considered dose-limiting. For patients with increased bilirubin, a total bilirubin > 4.8 nm / mL if direct bilirubin is > 0.3 mg / mL and indirect bilirubin is > 4.5 mg / mL, will be considered as dose limiting. Visible or visible KS lesions that, in the opinion of the Principal Investigator or Study Director, experience necrosis and / or bleeding as a result of a response to treatment with IL-12, will not constitute an adverse event or be considered a toxicity related to the drug. The above toxicities will be considered dose limiting unless, upon evaluation, it is found that the toxicity is probably or definitely due to the underlying condition of the patient. Patients with AIDS sometimes develop abnormal clinical or laboratory conditions in the absence of therapy (eg, fever from Pneumocystis carinii pneumonia (PCP).) For reasons of the protocol, patients develop dose-limiting toxicities that, after of evaluation, it is found that they are due to their AIDS, they will not be registered as such towards the definition of the highest tolerated dose.If said toxicity due to AIDS develops within the first 4 weeks of the administration of the drug or a patient chooses If you leave the study during this period, another patient may enter in your place.

Statistical Considerations The main objective of the study is to obtain information about the tolerability of EL-12 administered to patients with KS associated with HIV over a range of 5 increasing dose levels and to determine the highest tolerable dose. From 3 to 6 patients will enter each of the 5 dose levels, and up to 3 patients per dose level can be replaced as specified in the previous section of "Study Design." The highest tolerable dose will be defined as the highest dose level administered as part of the protocol where 0 of 6 or 1 of 6 patients experience dose-limiting toxicity. In determining the highest tolerated dose level, up to about 10 additional patients can be enrolled at this dose level in order to better define the tolerability and toxicity of this dose level and to preliminarily explore the activity of IL-12 in patients with KS associated with HIV. Therefore, the study would have a maximum accumulation of 55 patients. It is possible that, after reviewing the information from these or other studies of EL-12 administration, the accumulation of additional patients at a dose level below the highest tolerated dose can be substantiated. If this occurs, an amendment to the protocol will be made to allow additional patients to be enrolled at dose levels below the highest tolerated dose. Preliminary information will be obtained regarding the activity of EL-12 in patients with KS associated with HIV. As stated above, the evaluation of the response of the KS to an agent or regimen is difficult to assess by means of the commonly used oncological definitions. However, in an effort to standardize the evaluation of therapy against KS, the AIDS Clinical Trial Group Oncology Committee has devised a set of stage and response definitions for the KS. Therefore, we will use a modification of these criteria, as outlined in the previous section of "Response Criteria," to evaluate the response of patients with KS to the administration of IL-12. It should be noted that there is some variability of the observer in the evaluation of the number, size, nodularity and color of the lesions, and this should be taken into account when interpreting the measurements. In order to further explore the possible effect of EL-12 on the underlying HIV infection and immune function, evaluations of changes in viral load (testing for PCT of HIV RNA) will be made: the absolute number of cells CD4; Interferon-gamma levels of serum and / or urine, bFGF, VEGF, and EL-12; and subgroups TH1 / TH2 and production of EL- 12. To evaluate these changes, the main comparison will be between the results at week 12 and at entry. Parametric tests for comparison will not be used for exploratory purposes, and adjacent dose level groups can be combined to increase the power to see changes. Other time points (including week 4) will be examined and compared with the entry. Subjects of both genders and all racial / ethnic groups will be eligible for this study if they meet the eligibility criteria outlined in the previous section on "Eligibility and Enrollment Evaluation." To date, there is no information to suggest that differences in drug metabolism or response to disease would be expected in one group compared to another. Efforts will be made to extend the accumulation to a representative population, but in this preliminary study a balance must be found between considerations related to patient safety and limitations in the number of individuals exposed to potentially toxic and / or ineffective treatments, on the one hand , and the need to explore genres and ethnic aspects of clinical research, on the other hand. If differences are observed in the result that correlate with gender or ethnic identity, the accumulation can be expanded or a follow-up study can be written to investigate those differences more completely. As a practical matter, it should be noted that for reasons that are not completely understood, KS is relatively rare in females with HIV infection, and therefore it is expected that the vast majority of patients recruited in the study are males. The estimated annual accumulation for this test is 24 patients / year (2 patients / month). Therefore, if the protocol recruited the maximum number of patients (55), the estimated time to enroll these 55 patients would be ~ 2.5 years.

Research Ethics The nature of the research and the objectives of this trial, the procedures and treatments involved and their risks and disagreements of participation, potential benefits and potential alternative therapies will be carefully explained to the patient, and an informed consent document will be obtained. signed.

Information Report The NCI Cancer Treatment Evaluation Program (CTEP) will monitor this protocol and the information will be provided to the Clinical Triage Monitoring Service every 2 weeks using the Report Format of DCT Case or an acceptable magnetic tape format. The toxicities that occur must be reported to the CTEP as stipulated in Annex 1 of the CTEP Researchers Manual. Specifically, all toxicities (Grade 4) that threaten life and that may be due to drug administration, and all fatal events should be reported to the Investigational Drug Branch (telephone 230-2330, Fax. 230-0159) within 24 hours. A written report must be sent within 10 business days (ADE formats of the CTEP) to: Investigational Drug Branch P.O. Box 30012 Bethesda, MD 20824 The first occurrence of any new toxicity, regardless of the grade, must be reported to the drug monitor within the Investigational Drug Branch within 24 hours. An ADE format may be required. In addition, all ADEs will be reported to IRB, NCI within 10 business days.

Pharmaceutical Information IL-12 (NSSC # 672423) is a research agent manufactured by Genetics Institute. The IND will be maintained and the drug will be supplied by CTEP, DCTDC, NCI.

Formulation. This test will use human recombinant EL-12 (rhIL-12) as the product drug that will be supplied as a lyophilized powder in 5 mL bottles under a gentle vacuum. Each bottle will contain 50 μg of rhIL-12. The label attached to each vial of EL-12 will contain the appropriate information, including product name and quantity, batch number, storage conditions, sponsor's name, and appropriate regulatory precautions. The rhIL-12 bottles are intended for simple use only. Sterile water for injection (WFI) will be supplied to reconstitute the drug lyophilized product. WFI BACTERIOSTATIC MUST NOT BE USED TO RECONSTITUTE rhIL-12.

Storage. The drug product of lyophilized rhIL-12 must be stored in a refrigerated and safe facility at 2-8 ° C. The WFI can be stored at controlled room temperature. Dosing solutions are stable in the bottle after reconstitution for two (2) hours at 2 ° C to 8 ° C. Unit doses in the syringes can be maintained at controlled room temperature and should be used within four (4) hours after preparation.

Preparation. The drug product of rhIL-12 will be supplied as a lyophilized powder in a 5 mL bottle containing 50 μg of rhIL-12. The dose of the study drug will be calculated using the weight in kilograms (kg) of the subject. Patients will be weighed at each clinic visit and a new dose will be calculated to determine the dose of study drug that will be given if the patient's weight has changed ± 5% from the benchmark. The drug lyophilized product will be reconstituted with either 5 mL or 1 mL of sterile WFI. Reconstitution with 5 mL of WFI will provide a dosage solution of 10 μg / mL to be used for the dose level of 100 ng / kg. Reconstitution with 1 mL of WFI will provide a 50 μg / mL dosing solution to be used for the 300 ng / kg dose level and all higher dose levels. To reconstitute the rhIL-12, the WFI is injected into the bottle through the cap. It may be necessary to remove some of the air from the bottle to facilitate the dilution of the 5 mL. For the dilution of 5 mL, it is recommended that approximately 2 mL of WFI be added to the powder vial first. Some air should be removed with the SAME syringe before adding another 3 mL. The bottle in then gently rolled to dissolve the powder. Reconstitution will be complete in approximately one (1) minute. No flask with discoloration or particulate material should be used after reconstitution. The following tables provide examples of dose volumes based on subject weight ranging from 40 to 100 kg for planned study dose levels of 100 ng / kg and all levels >; 300 ng / kg using one of the two dosing solutions. The table specifies the dosage solution (10 or 50 μg / mL) to be used for the preparation of study drug doses.

Sample calculation for a patient of 65.4 kg to receive the dose level of 300 ng / kg: Reconstitution with 1 mL of WFI will provide dosing solution of 50 μg / mL = 50,000 ng / mL Total dose = 65.4kx x 300 ng / kg = 19,620 ng Volume injected = Total dose - concentration = 19,620 ng • * - 50,000 ng / ml = 0.39 mL Administration Procedures. The dosage will be calculated on the basis of the actual body weight in kg as indicated by the previous section of "Drug Administration." The study drug will be administered by subcutaneous injection (SC). No more than 2 mL should be injected subcutaneously as a single injection.

Accountability of the Drug and Drug Application. The drug can be requested by completing a Clinical Drug Application (NIH-986) and mailing it to the Drug Management and Authorization Section, DCTDC, NCI, EPN Room 707, Bethesda, MD 20892 or by faxing it to (301) 480-4612 . For questions call (301) 496-5725.

Registration of Drug Inventories. The investigator or responsible party designated by the investigator must maintain a careful record of the inventories and disposition of all drugs received from the CDTDC, using the NCI Drug Accountability Record format. (See NCI Investigators Handbook for Procedures for Drug Accountability and Storage).

Example 2 - Results of Clinical Treatment of KS Associated with AIDS, with IL-12 Fifteen (15) patients were enrolled in relation to the protocol delineated in Example 1. Patients were treated with 100 ng / kg (5 patients), 300 ng / kg (6 patients) and 500 ng / kg (4 patients) per protocol. The mean CD4 count for enrolled patients was 201 cells / mm3 (range 17-602 cells / mm3). The average viral load for enrolled patients was 2384 copies / mL (range 0-156,996 copies / mL). The stage of progression of the KS in the enrolled patients was recorded using the TIS system of ACTG stages as follows: Good progression 0 patients Poor prognosis 15 patients Ti 12 patients Ii 7 patients If 11 patients Enrolled patients who had received prior treatment as follows: None 1 patients Chemotherapy 9 patients Ehterferon 3 patients Local Therapy 2 patients Experimental Therapy 10 patients The treated patients experienced the following laboratory toxicities (as defined in the protocol of Example 1): Toxicity Grade 2 Grade 3 Grade 4 Leukopenia 8 3 0 Neutropenia Not assessed 6 4 SGPT 0 3 0 SGOT 3 2 0 T. Bilirubin 4 1 0 Glucose 4 0 0 Four (4) patients received G-CSF; three continued with EL-12 without additional neutropenia, while the fourth was intolerant to G-CSF and unable to continue with IL-12 therapy.

The treated patients presented the following clinical toxicities (as defined in the protocol of Example 1): - Fever syndrome, sweating, fatigue, headache - Beginning with the initiation of therapy - Spontaneous resolution in 1 to 2 weeks of the continued therapy -Injection site -Keep-minimum clearance From the submission of this application, the treated patients had shown the following clinical results: -Dose level of 100 ng / kg: 5 patients -1 presumed toxoplasmosis CNS week 1 -3 progression weeks 2, 4 and 4 - 1 stable disease completed 26 weeks of therapy -Level of 300 ng / kg: 6 patients -1 liver toxicity dose-limiting week 3 -2 partial responses -1 neutropenia with G-CSF intolerance week 22 -1 continues in week 13+ -3 continues stable in weeks 25+, 19+ and 15+ -Level of 500 ng / kg: 4 patients -4 continues stable in weeks 9+, 8+, 7+ and 4+ These data demonstrated that IL-12 is well tolerated in patients with KS and that treatment with IL-12 produces a clinical response in response to treatment of KS.

References Mosmann TR, Cherwinski H, Bond MW, Giedlin MA, Coffman RL. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins (Two types of murine T helper cell clones I. Definition according to profiles of activities of lymphokine and secreted proteins.) J Immunol 1986; 136: 2348-57. Scott P, Kaufman SHE. The role of T-cell subsets and cytokines in the regulation of infection (The role of subgroups of T cells and cytokines in the regulation of infection). Immunol. Today 1991; 12: 346-8. Sher A, Gazzinelli RT, Oswald IP, et al. Role of T-cell derived cytokines in the downregulation of immune responses in parasitic and retroviral infection (Role of cytokines derived from T cells in the hyporegulation of immune responses in parasitic and retroviral infections). Immunol. Reviews 1992. 127: 183-204. Romagnani S. Human TH1 and TH2 subsets: doubt no more (Human TH1 and TH2 subgroups: no more doubts). Immunol Today 199 1; 12: 256-7. Clerici M, Hakim FT, Venzon DJ, et al. Changes in interleukin-2 and interleukin-4 production in asymptomatic, human immunodeficiency virus-seropositive individuáis (Changes in the production of interleukin 2 and interleukin 4 in seropositive individuals infected with the human immunodeficiency virus, asymptomatic). J Clin Invest 1993; 91: 759-65. Maggi E, Mazzetti M, Ravina A, et al. Ability of HIV to promote THL to THO shift and to replicate preferentially in TH2 and THO cells [see comments] (Ability of HIV to promote a change from THI to THO and to replicate preferentially in TH2 and THO cells [see comments]). Science 1994; 265: 244-8. Trinchieri G. Interleukin-12: a cytokine produced by antigen-presenting cells with immunoregulatory functions in the generation of T-helper cells type 1 and cytotoxic lymphocytes (Interleukin 12: a cytokine produced by antigen-presenting cells with immunoregulatory functions in the generation of type 1 T helper cells and cytotoxic lymphocytes). Blood 1994; 84: 4008-27.

. Brunda MJ, Luistro L, Warrier RR, et al. Antitumor and antimetastatic activity of interleukin 12 against murine tumors (Antitumor and antimetastatic activity of interleukin 12 against murine tumors). J Exp Med 1993, 178: 1223-30. . Scott P. IL-12: initiation cvtokine for cell-mediated inunit (IL-12: initiation cytokine for cell-mediated immunity). Science 1993; 60: 496-7. 10. Clerici M, Lucey DR, Berzofsky JA, et al. Restoration of HFV-specific cell-mediated immune responses by interleukin-12 in vitro (Restoration of immune responses mediated by HIV-specific cells by interleukin 12 in vitro). Science 1993; 262: 1721-4. 11. Voest EE, Kenyon BM, MS OR, Truitt G, RJ DA, Folkman J. Inhibition of angiogenesis in vivo by interleukin 12 (Inhibition of angiogenesis in vivo by interleukin 12). J Nati Cancer Inst 1995; 87: 581-6. 12. Angiolillo AL, Sgadari C, Taub DD, et al. Human interferon-inducible protein 10 is a potent inhibitor of angiogenesis in vivo (Interferon-inducible human protein 10 is a potent inhibitor of angiogenesis in vivo). J Exp Med 1995; 182: 155-62. 13. Foli A, Saville MW, Baseler MW, Yarchoan R. Effeets of the Thi and Th2 stimulatory cytokines interleukin-12 and interleukin-4 on human immunodeficiency and virus replication (Effects of Thi and Th2 stimulatory cytokines, interleukin 12 and interleukin 4 , in the replication of the human immunodeficiency virus). Blood 1995; 85: 2114-2i.

Table 1 COMMON TOXICITY CRITERIA GRADE TOXICITY 1 2 WBC = 4.0 3.0-3.9 2.0-2.9 1.0 - 1.9 < 1.0 PLT WNL 750 - normal 50-0 - 74.9 25.0-49.9 < 25.0 Hgb WNL 100 -normal 8.0-10.0 6.5-7..9 < 6.5 Bands of = 2.0 1.5-1.9 1.0-1.4 0.5-0.9 < 0.5 Granulocytes Lymphocytes = 2.0 1.5-1.9 1.0-1.4 0.5-0.9 < 0.5 Hemorrhage (clinical) none Soft, no large transfusion, 1-2 units large, 3-4 units Massive < 4 units of transf. per episode transf. per episode transf. per episode Infection none Mild Moderate severe threat life Nausea None Able to eat intake significantly no reasonably diminished intake, but may be important to eat Vomiting none 1 episode in 24 hours 2-5 episodes in 24 hours 6-10 episodes in 24 < 10 episodes in 24 hrs, or need parenteral support Diarrhea no Increment of 2-3 Increase of 4-6 Increase of 7-9 Increase of = 10 stools / day before stools / day, or stools / day, or stools / day, or Rx nocturnal stools, or incontinence, or severe bloody diarrhea, or moderate cramping cramps requires parenteral support Stomatitis no sores without pain, erythema, painful erythema, erythema edema painful, edema requires support or mild ulceration or ulcers but can or ulcers but can parenteral or enteric eat eat Bilirubin WNL • • < 1.5xN 1.5- 3.0 xN > 3.0xN Transaminase WNL = 2.5xN 2.6- 5.0 xN 5.1- 20.0 xN > 20.0 x N (SGOT, SGPT) Fosf. Ale or WNL = 2.5xN 2.6- 5.0 xN 5.1- 20.0 xN > 20.0 x N 5 'Nucleotidase Liver (clinical) No change with respect to - • - Precoma hepatic coma reference Creatinine WNL < 1.5xN 1.5- 3.0 xN 3.1- 6.0 xN > 6.0xN Proteinuria without change 1 + or 2-3 + 0 4+ or nephrotic syndrome 0.3 go 0.3 - 1.0 g or > 1.0go < 3g? 3 -10 gil > 10flile Negative hematuria only micro Large, no large clots + clots requires transfusion Alopecia without hair loss Scarce loss of hair Major or total loss •• of hair Pulmonary null or no change Asymptomatic, with dyspnoea in exercise dyspnea at a normal level dyspnea at rest abnormality in PET's important activity Cardiac dysrhythmias none Asymptomatic, Recurrent or persistent , requires treatment requires monitoring or transient without requiring without requiring hypotension therapy, or tachycardia ventricular therapy or fibrillation Cardiac function none Asymptomatic, decreases Asymptomatic, decreases mild CHF in response to severe or refractory CHF at rest, traction at rest, ejection therapy traction in less than ejection by more than 20% 20% of the reference value reference value Ischemia No specific cardiac symptoms Asymptomatic, changes of angina without evidence of acute infarction of wave flattening ST and T waves suggest myocardial infarction T ischemia Table 1 (Continued) COMMON TOXICITY CRITERIA GRADE TOXICITY 2 Pericardial cardiac Null Asymptomatic, effusion, Pericarditis (symptomatic pain, effusion, tamponade, requires no intervention chest affection, changes of drainage requires urgent drainage ECG) Null or no change hypertension Asymptomatic, increase Recurrent or increase requires therapy transient hypertensive crisis in more than 20 persistent in more than 20 m Hg (D) or to mm Hg (D) or to > 150/100 if WNL > 150/100 if previous WNL, no prior requirement, treatment is required treatment No or no change hypotension Changes that do not require Requires replacement of therapy and requires therapy and therapy (including fluids or other therapy, hospitalization, hospitalization by> 48 orthopathic transient) but not hospitalization resolved within 48 hrs of the hrs after suspension of agent agent suspension Neuro-sensitive null or no change Minor paresthesias, loss of mild or moderate loss, severe loss of tendon reflexes of objective sensitivity; Objective sensitivity or deep paresthesias moderate paresthesia that interferes with Neurotrophic function null or no change Subjective weakness; no mild objective weakness without objective weakness with paralysis objective findings major damage to function impairment Neuro-cortical function null Mild drowsiness or drowsiness or agitation severe drowsiness, coma, access, moderate agitation agitation, confusion, toxic psychosis disorientation or hallucinations Neuro- cerebellar null Slight uncoordination, intentional tremor, locomotor ataxia Cerebellar necrosis Disdiadochinesis dysmetria, cut language, pistag or Neuro-emotional without change Mild anxiety or depression moderate anxiety or severe anxiety o Suicidal ideation depression depression Neuro-pain of null Mild moderate or severe but no cede and severe transient head Neuro-constriction null or no change Mild Moderate Severe ileus > 96 hrs Neuro-hearing No or no change Attic asythus, loss of Tinnitus hearing loss deafness not susceptible to hearing only in interfering with correcting audiometry function but susceptible to correction with hearing aid Neuro-vision Null or no change loss of vision blindness symptomatic subtotal Skin Null or unchanged Macular or macular or macular rash, papular or exfoliative or papular dermatitis or papular dispersed or vesicular symptomatic and ulcerative dermatitis erythema that is erythema with pruritus or asymptomatic generalized other associated eruption symptoms Transient null rash allergy, Urticaria fever, serum sickness fever, anaphylaxis drug < 38 ° C, 100.4 drug - 38c. 100.4F bronchospasm, requires ° F mild bronchospasm of parenteral measures Fever with no absence 37.1 -38.0 c 38.1 -40.0c > 40.0c > 40.0c (104.0F) for 98.7- 100.4F 100.5 - 104.0F > 104.0F more than 24 hrs or infection fever for less than 24 hrs. accompanied by hypotension Local null Pain pain and swelling with ulceration plastic surgery indicated inflammation or phlebitis Table 1 (Continued) CO TOXICITY CRITERIA GRADE TOXICITY 0 1 2 3 4 Gain / loss of weight < 5.0S 5.0-9.9% 10.0 - 19.9% > 20.0% Hyperglycemia < 116 116-160 161-250 251-500 > 500 or ketoacidosis Hypoglycemia > 64 55-64 40-54 30-39 < 30 Amylase WNL < 1.5xN -1.5- 2.0 xN 2.1- 5.0 xN > 5.1xN Hypercalcemia < 10.6 10.6-11.5 11.6-12.5 12.6-13.5 = 13.5 Hypocalcemia > 8.4 8.4-7.8 7.7-7.0 6.9-6.1 < 6.0 Hypomagpesemia > 1.4 1.4-1.2 1.1-0.9 0.8-0.6 = 0.5 Fibrinogen WNL 0.99- 0.75 xN 0.74- 0.50 XN 0.49- 0.25 xN < 0.24 N Prothrombin time WNL 1.01- 1.25 xN 1.26- 1.50 xN 1.51- 2.00 xN > 2.00xN WNL Time 1.01- 1.66 N 1.67- 2.33 xN 2.34- 3.00 xN > 3.00xN partial thromboplastin COMPLEMENTARY TOXICITY CRITERIA FOR BLOOD PLUGGOTTEN CELL SUPPORT OR AUTOMOTIVE BONE MARROW SUPPORT STUDIES Grade 5 Death due to bacterial or fungal infection or neutrophil-associated hemorrhage < 500 / ul or platelets < 10,000 / ul more than 8 weeks after the bone marrow transplant. Grade 4 Neutrophils < 500 / ul and / or platelets < 10,000 / ul for a duration above the 8 weeks. Grade 3 Neutrophils < 500 / ul and / or platelets < 10,000 / ul for a duration of 4-8 weeks. Grade 2 Neutrophils < 500 / ul and / or platelets < 10,000 / ul for a duration of up to 4 weeks. Grade 1 Neutropenia and / or thrombocytopenia, but never neutrophils < 500 / ul and the platelets never < 10,000 / ul. All other non-hematological toxicities should be graded by the Co Toxicity Criteria.

Claims (14)

1. Novelty of the Invention 1. A method for the treatment of Kaposi's sarcoma in a mammalian subject, said method comprising administering to said subject a therapeutically effective amount of IL-12 or a biologically active fragment or subunit thereof.
2. 2. The method of claim 1, wherein said subject has AIDS.
3. 3. The method of claim 1, wherein said IL-12 is administered as a protein.
4. 4. The method of claim 1, wherein said IL-12 is administered in the form of DNA encoding IL-12.
5. The method of claim 3, wherein said protein is administered in a dose of from 100 to 1000 ng / kg body weight of the subject.
6. The method of claim 5, wherein said protein is administered at a dose selected from the group consisting of 300 ng / kg of subject's body weight, 500 ng / kg of subject's body weight, 625 ng / kg of body weight of the subject and 750 ng / kg of body weight of the subject.
7. The method of claim 6, wherein said protein is administered in a dose of 300 ng / kg of body weight of the subject.
8. 8. The method of claim 9, wherein said subject has AIDS.
9. 9. A method for the inhibition of angiogenesis in a lesion associated with Kaposi's sarcoma in a mammalian subject, said method comprising administering to said subject a therapeutically effective amount of IL-12 or a biologically active fragment or subunit thereof. .
10. The method of claim 7, wherein said IL-12 is administered as a protein.
11. The method of claim 10, wherein said IL-12 is administered in the form of DNA encoding IL-12
12. 12. The method of claim 11, wherein said protein is administered in a dose of from 100 to 1000 ng / kg body weight of the subject.
13. 13. The method of claim 12, wherein said protein is administered in a dose selected from the group consisting of 300 ng / kg body weight of the subject, 500 ng / kg body weight of the subject, 625 ng / kg body weight of the subject and 750 ng / kg of subject's body weight. The method of claim 13, wherein said protein is administered in a dose of 300 ng / kg of body weight of the subject. Extract of the Description Methods for using IL-12 to treat Kaposi's sarcoma (KS), particularly KS associated with AIDS are described.