Pharmaceutical compositions containing giroxina and phospholipase A.sub.2 and methods of increasing a patient's CD.sub.4 count using the pharmaceutical compositions

Pharmaceutical compositions for treatment of humans suffering from the symptoms of acquired immune deficiency syndrome (AIDS) or its precursors, lymphadenopathy syndrome (LAS) or AIDS-related complex (ARC) or secondary diseases relating thereto and other immunodeficiency pathologies or tumors resulting exclusively from immune deficiency and others such as H.I.V. The compositions contain a therapeutically active amount of phospholipase A.sub.2 and giroxina, preferably where the phospholipase A.sub.2 is isolated from the venom of Crotalus durissus terrificus or form the venom of Micrurus frontalis altirostris and the giroxina is isolated from the venom of Crotalus durissus terrificus. Also disclosed is the process for preparing such pharmaceutical composition and the process for treating patients suffering from the aforementioned afflictions.

INTRODUCTION AND BACKGROUND 
The present invention relates to pharmaceutical compositions having active 
ingredients derived and isolated from the venom of certain snakes for 
treatment of humans suffering from the symptoms of acquired immune 
deficiency syndrome (AIDS), or its precursors, lymphadenopathy syndrome 
(LAS) or AIDS-related complex (ARC) or secondary diseases relating 
thereto, and other immunodeficiency pathologies or tumors resulting 
exclusively from immune deficiency and others, such as those caused by 
H.I.V. The present invention also relates to a process for the preparation 
of such pharmaceutical compositions and also to a process for the 
treatment of humans suffering from the above diseases by administering to 
such humans a therapeutically effective amount of such pharmaceutical 
compositions wherein the active ingredients are isolated from snake venom. 
The biological activity of certain snake venoms has been recognized in the 
art and is known. Research to determine the composition of and the 
specific effect produced by the components of snake venoms has been 
ongoing. It has been found that snake venom contains a plurality of 
proteinaceous and other components or substances. Certain snake venoms or 
fractions thereof have exhibited therapeutical activity as anticoagulants 
and as analgesics. 
U.S. Pat. No. 3,657,416 discloses an enzyme isolated from the venom of the 
snake Agkistrodon rhodostoma and the use of this enzyme for the 
therapeutic treatment of humans. According to U.S. Pat. No.3,888,977, 
detoxified and neurotropically active modified snake venom neurotoxins 
(derived from a species of the Bungarus genus) are used for the treatment 
of progressive degenerative diseases of the nervous system which involve 
the function of motor nerve cells from the origin of such cells to the 
neuromuscular junction, as well as elements of the central nervous system 
including axons, nerve myelin sheaths, etc.; such diseases include 
amyotrophic lateral sclerosis, multiple sclerosis, kuru, polymyositis, 
certain meningitides, muscular dystrophy, and the like. Detoxification of 
the venom is performed by treatment with formaldehyde, fluorescein dyes, 
ultraviolet light or gentle oxygenation at relatively low temperatures. 
U.S. Pat. No. 3,888,977 relates to a modified neurotoxic snake venom 
detoxified e.g. by oxygenation to form an atoxic, neurotropically active 
therapeutical composition. The venom must be at least in part derived from 
the genus Bungarus, but preferably also contains venoms of the genus Naja 
or Crotalus durissus terrificus. The composition mitigates the progress of 
degenerative neurological diseases by blocking nerve cell receptors. 
U.S. Pat. No. 3,888,977 is, however, silent in disclosing the chemical 
components or composition of the modified venom. It is stated in U.S. Pat. 
No. 3,888,977 that the venom of snakes contains a multitude of chemical 
compounds including various enzymes. The exact function of the enzymes in 
the venom is not understood, according to the patent at column 3, and, 
indeed, the enzymes may not have any direct function in the toxic effect 
of venom. The enzymes may have other functions beneficial to the snake in 
utilizing the victim of its bite as food. 
In U.S. Pat. No. 4,126,676, similar methods of treatment of neurological 
disorders are disclosed and detoxified, modified neurotoxins derived from 
the Naja genus of snake are used. 
U.S. Pat. No. 4,341,762 discloses compositions having pharmacological 
activity and which contain, in an administrable form, at least one 
post-synaptic neurotoxin, at least one pre-synaptic neurotoxin, and at 
least one component capable of stimulating the immune mechanisms of the 
body. The post-synaptic neurotoxin component preferably contains the 
.alpha.-toxin obtained from the venom of an elapid snake belonging to a 
species of the genera Naja, Ophiophagus or Dendroaspis. The pre-synaptic 
neurotoxin component preferably contains .beta.-bungarotoxin obtained from 
the venom of the elapid snakes Bungarus multicinctus and the component 
capable of stimulating the immune mechanisms of the body preferably 
contains a venom obtained from a viperid snake, particularly a venom 
obtained from a snake belonging to the family Vipera, subfamily 
Crotalinae. 
The compositions disclosed in U.S. Pat. No. 4,341,762 may include the whole 
venom from which a particular activity is being sought or only a 
particular fraction or fractions of the venom. Thus, the pre-synaptic 
neurotoxin may consist of the whole venom of a Bungarus species. A whole 
venom from the Naja species contains post-synaptic neurotoxins which can 
potentially complement the pre-synaptic activity of the Bungarus component 
of the mixture. The component capable of stimulating the immune mechanisms 
of the body, the viperid component, is preferably used and is present as 
the whole venom since such venoms contain a plurality of enzymatic 
substances. 
The compositions disclosed in U.S. Pat. No. 4,341,762 stimulate the 
production of the substance known as "interferon" or a precursor thereof. 
This stimulation is due to the projected presence of proteins having at 
least portions which resemble double-strand RNA molecular structures or 
which contain enzymatic activities capable of converting precursor 
proteins to interferon stimulating or activating agents. The compositions 
set forth in U.S. Pat. No. 4,341,762 are useful as antiviral and 
antiautoimmune agents by the stimulation of the part of the body's immune 
system which involves interferon or the complex activity within the body 
attributed to interferon. The compositions disclosed in U.S. Pat. No. 
4,341,762 can be used in the treatment of the symptoms of poliomyletis, 
herpes simplex, herpes zoster, herpes genitalis, and diseases such as 
multiple sclerosis and amyotrophic lateral sclerosis and degenerative 
neurological disorders. The mixture of the above venoms and/or venom 
fractions is preferably finished in the form of solutions formed with 
saline solution preserved with THIMEROSAL.RTM. and are administered by 
injecting into the body a sterile pyrogen-free solution. The chemical 
structure of the composition of the components of the snake venoms or 
fractions thereof are not identified nor disclosed in U.S. Pat. No. 
4,341,762. 
The compositions set forth in U.S. Pat. No. 4,741,902 are an improvement of 
those disclosed in U.S. Pat. No. 4,341,762. These compositions also 
contain effective amounts of at least one post-synaptic neurotoxin and at 
least one pre-synaptic neurotoxin, but the component capable of 
stimulating the immune mechanisms of the body is the viperid "b" fraction 
obtained from elution of the viperid venom on SEPHADEX.RTM. G-50 column in 
place of the whole viperid snake venom. The composition is allegedly 
useful in the treatment of the diseases enumerated in U.S. Pat. No. 
4,341,762. The venom fraction obtained after elution on a SEPHADEX.RTM. 
G-50 column is free of certain enzymes such as L-amino acid oxidase and 
phosphodiesterase, and this imparts particular utility to the composition 
due to the absence of a hemorrhagic effect when used in the treatment of 
mammals 
U.S. Pat. No. 5,002,766 is directed to a method of treating a patient 
having acquired immune deficiency syndrome (AIDS) or its precursors, 
lymphadenopathy syndrome (LAS) or AIDS-related complex (ARC), by 
administering a composition comprising trypsin, .alpha.-chymotrypsin, 
papain, pancreatin, bromeline, lipase, amylase and rutin. According to the 
disclosure of the patent, the above catabolic enzymes show unexpected 
successes in the improvement of the condition of the patients suffering 
from AIDS or precursors or secondary diseases related thereto. 
It is known that acquired immune deficiency syndrome (AIDS) and its 
precursors are caused by a retrovirus called human immunodeficiency virus 
(HIV). Infection with HIV disturbs the whole immunodefense mechanism of 
the human organism, first of all by affecting the key positions of 
immunodefense, namely the helper cells of the T system. Such T4 (helper) 
cells affected by the infection with HIV are no longer capable of 
performing their central role in the regulation of the immunoresponse. The 
disturbing effect caused by infection with HIV is the result of a direct 
or indirect reduction of the T4 (helper) cells. 
Infections with HIV result in disturbances of the equilibrium between the 
different T cell populations. Thus, the balance between the T4 (helper) 
and T8 (suppressor) cells is changed and is shifted in favor to the T8 
cells at the expense of the T4 cells. As a result, the T4/T8 ratio, being 
generally about 1.4 to 2, is shifted to a T4/T8 ratio of 1:2 or even lower 
values. The HIV-induced reduction of the T4 cells allows pathogenic 
organisms (e.g., viruses, bacteria, fungi or protozoans), which are 
harmless in case of a normal balance between the different T cell 
populations, to meet with favorable conditions in patients having the 
above disproportion between T4 and T8 cells. Persons suffering from AIDS 
may be subject to attack by different pathogenic organisms and 
carcinomatous degenerations of tissue (e.g., Kaposi sarcoma) and also may 
be subject to attack by other malignant diseases. 
AIDS is one of the most serious menaces to mankind. Although various drugs 
and treatment have been developed, none of them provides any significant 
success against the affliction. 
SUMMARY OF THE INVENTION 
It is an object of the present invention to provide a pharmaceutical 
composition which mitigates the symptoms of AIDS or its precursors (LAS or 
ARC) or secondary diseases related thereto or other 
immune-deficiency-pathologies or tumors resulting exclusively from immune 
deficiency and others such as HIV, and keeps patients suffering from the 
above diseases symptom-free for a longer period of time. The main field of 
application of the composition according to the present invention is the 
treatment of patients suffering from the symptoms of AIDS. 
It is a further object of the present invention to provide a method of 
treatment which mitigates the symptoms of AIDS or its precursors (LAS or 
ARC) or secondary diseases related thereto or other 
immune-deficiency-pathologies or tumors resulting exclusively from immune 
deficiency and others such as HIV, and keeps patients suffering from the 
above diseases symptom-free for a longer period of time. 
According to an aspect of the present invention, there is provided a 
pharmaceutical composition for treatment of humans suffering from acquired 
immune deficiency syndrome (AIDS) or its precursors, lymphadenopathy 
syndrome (LAS), or AIDS-related complex (ARC), or secondary diseases 
related thereto or other immunodeficiency-pathologies or tumors resulting 
exclusively from immune deficiency and others such as HIV, comprising a 
therapeutically active amount of phospholipase A.sub.2 and giroxina in 
admixture with suitable inert pharmaceutical carriers or diluents. 
According to a further aspect of the present invention, there is provided a 
process for the preparation of a pharmaceutical composition for the 
treatment of humans suffering from acquired immune deficiency syndrome 
(AIDS), or its precursors lymphadenopathy syndrome (LAS) or AIDS-related 
complex (ARC), or a secondary disease related thereto, or other 
immunodeficiency-pathologies or tumors resulting exclusively from immune 
deficiency and others such as HIV, comprising admixing a therapeutically 
active amount of phospholipase A.sub.2 and giroxina with suitable inert 
pharmaceutical carriers or diluents. 
According to a still further aspect of the present invention, there is 
provided a method of treatment of a patient suffering from acquired immune 
deficiency syndrome (AIDS), or its precursors, lymphadenopathy syndrome 
(LAS) or AIDS-related complex (ARC), or secondary diseases related thereto 
or other immunodeficiency-pathologies or tumors, resulting exclusively 
from immune deficiency and others such as HIV, comprising administering to 
the patient in need thereof a composition comprising a therapeutically 
active amount or phospholipase A.sub.2 and giroxina in admixture with an 
inert pharmaceutical carrier or diluent.

DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION 
The phospholipase A.sub.2 component of the composition according to the 
present invention is an enzyme (Enzyme Code 3.1.1.4) which is also 
denominated as phosphatydil 2-acyl hydrolase or Lecitinase A. 
Phospholipase A.sub.2 may be isolated from the dried poison of Crotalus 
durissus terrificus (cascabel of Argentina). It is designated in the 
International Catalogue Sigma Chemical Company, Biochemical Organic 
Compounds (for Investigation and Diagnostic Reagents), U.S.A. and Canada, 
at page 812 of Edition 1990 and at page 841 of Edition 1991 as "Product 
No. 5910". Phospholipase A.sub.2 appears as a lyophilized dust containing 
50% of proteins (standardized by the Biset method) and has an activity of 
200 U.V. times per mg., at pH 8.9 and 25.degree. C., using as substrate 
L-A Phosphatydil choline of soya. Phospholipase A.sub.2 can also be 
isolated from the dried poison of Micrurus frontalis altirostris, a coral 
snake of Argentina. 
The giroxina component of the composition according to the present 
invention is a basic polypeptide, molecular weight 3000 daltons, pH 10 
containing four sulfur bridges. Giroxina was first disclosed to be 
contained in the so-called "Crotoxina Complex" (Slotta and Frankel-Conrat: 
Chemical Studies on Ofidios Venoms (1038-39), Accounts of the Butantan 
Institute, Volume 12, pages 505-513). Crotoxina complex is described as 
having a molecular weight of 33,000 daltons. Later it turned out that the 
basic component Crotoxina actually contains was "Crotamina", a basic 
polypeptide, pH 10.3, molecular weight 5430 daltons, containing four 
sulfur bridges. 
Crotoxina is also disclosed to be a complex containing an acidic protein 
(pH 3.7, molecular weight 9000 daltons) and a basic: protein (pH 8.65, 
molecular weight 12,000 daltons). The total molecular weight of Crotoxina 
is set forth to be 21,000 daltons. 
Alexander VRS Voet described in his book entitled "The Why, When and How of 
Ophibians (snakes)", published in 1985 by the Editorial American Lee 
Publishing House (Argentina), on page 423, that giroxina is a basic 
polypeptide with a molecular weight of 3,000 daltons, pH 10, and having 
four sulfur bridges. It was disclosed, however, to be just a minimal 
fraction of the complete venom complex and no pharmaceutical utility of 
giroxina is set forth in the book. 
Giroxina can be isolated from the dried snake venom of Crotalus durissus 
terrificus. 
Phospholipase A.sub.2 and giroxina can be readily isolated from the dried 
venom of Crotalus durissus terrificus. The isolation of these components 
and the separation thereof from other enzymes, proteins and polypeptides 
present in the venom. (many of which are toxic) is based on the thermal 
stability of phospholipase A.sub.2 and giroxina. 
The fresh venom Crotalus durissus terrificus obtained by direct extraction 
is desiccated with the aid of a vacuum pump and/or in the presence of a 
dehydrating agent (e.g., calcium chloride). Desiccation is carried out at 
a temperature at which the majority of the other components (enzymes, 
neurotoxins, etc.) present in the fresh venom are destroyed by 
decomposition. A preferable temperature is 65.degree. C. The desiccated 
residue is a crystalline or pseudocrystalline powder. 
The weight ratio between phospholipase A.sub.2 and giroxina is within the 
range of 20:1 and 4:1, preferably between 8:1 and 9:1. According to a 
particularly preferred embodiment of the present invention, the weight 
ratio of phospholipase A.sub.2 and giroxina is about 8.1:1. 
The pharmaceutical compositions of the present invention may contain any 
suitable pharmaceutical carriers or diluents known to the art. It has been 
found to be advantageous to use a sodium chloride solution as diluent. The 
concentration of the sodium chloride solution may be preferably about 0.17 
mole/liter. The sodium chloride solution may preferably contain a 
conservator. Any suitable conservator (e.g., THIMEROSAL.RTM.) being inert 
to the active ingredients may be used. 
The pharmaceutical compositions according the present invention may 
optionally contain further components which strengthen the activity of the 
composition. For this purpose tioctic acid, magnesium lactate and/or 
B.sub.6 vitamin may be used. 
The composition according to the present invention may be preferably 
prepared by dissolving phospholipase A.sub.2 and giroxina in the 
weight-ratio disclosed above in a sodium chloride solution which contains 
a conservator. Dissolving is performed at a suitable temperature to 
dissolve the desired components, preferably at 76.degree. C. The solution 
is stirred as the components enter into solution. The solution thus 
obtained is placed into vials and the vials are hermetically sealed. 
The composition according to the present invention is preferably 
administered subcutaneously, though other routes of administration are 
possible (e.g., sub-lingual). The daily dosage depends on the seriousness 
of the disease and of the condition of the patient and is not related to 
the patient's weight. A preferred daily dose amounts to 0.10-0.30 ml of 
the solution prepared according to Example 1, infra. It is recommended to 
administer 0.15-0.18 ml as maintenance dose to patients who have recovered 
their normal weight. 0.10 ml would be administered to those patients 
having CD4 counts of &gt;700 and &lt;1200. On starting treatment of patients 
highly affected by AIDS virus, it is expedient to use a dose of 0.3 ml for 
at least 30 days ("highly affected" is defined as those AIDS patients to 
are in an advanced state of sickness or with advanced symptoms of 
sickness, technically measurable by the quantity of CD4 lymphocytes (i.e., 
less than 700 CD4 lymphocytes, which is below the normal range of 
1200-1400)); the dosage may be reduced by 0.05 ml per month as the CD4 
count increases. In more serious cases double (200%) of the above amounts 
may be administered. The treatment is to be carried out dally at least for 
60-90 days, and the patient's condition is checked every 20-25 days by 
means of clinical and laboratory controls. If necessary the daily 
treatment may be continued up to a total period of 120 days or more (days 
91-120). 
The pharmaceutical composition of the present invention when kept in a 
refrigerator at a temperature of +4.degree. C. maintains its biological 
activity for at least 120 days. Freezing alters the active ingredients and 
therefore the composition should not be subjected to freezing. 
Without being bound by theory, it has been found that the composition of 
the present invention does not act on the virus or tumor but unexpectedly 
induces an energic hypothalamic stimulation and actually causes a 
"hypothalamic shock" which results in a feeling of well-being (e.g., 
disappearance or decrease in pain, discomfort, adenopathies, loss of 
weight, and depression) and an increase of the immunoindividual-response 
(i.e. number of CD4 lymphocytes), allowing an excellent response with 
great and successful potential. In contrast to other drugs used in the 
treatment of AIDS, the composition according to the present invention does 
not act directly against the viruses or on the natural operators of the 
immune-lymphocytes-response (macrophagues, histiocytes, etc.), but exerts 
its effect on the hypothalamus - - - i.e., on the "orchestra director" of 
the immune system - - - making the individual able to defend himself from 
the viral attack, until the virus' pathogenic capacity is exhausted. 
A further advantage of the composition of the present invention over drugs 
hitherto used in the treatment of AIDS (e.g., AZT, interferon, Timoduline) 
is that the favorable effects of the inventive composition are not 
followed by further attacks which can be even more serious for the 
immunity deficiency. The effect on the individual of the composition 
according to the present invention is continuous, often lasting several 
months (between 90 and 120 days) after the completion of the treatment. 
A further significant advantage of the composition of the present invention 
resides in the absence of serious side-effects, which are characteristic 
of the hitherto known drugs, especially of AZT. The pharmaceutical 
composition of the present invention may only cause a light rubefaction 
and temporary irritations in the areas of application if it is used in the 
above recommended doses or in doses up to 100 or 200% of the suggested 
amount, which increased doses may be necessary in some cases. No serious 
secondary effects or side-effects were observed. 
Since the composition of the present invention can not be applied 
simultaneously with cortisone or similar drugs, it is a non-aggressive 
therapy and thus the effects of aggressive therapies (e.g., surgical, 
chemotherapy, etc.) can be diminished. The inventive composition is 
perfectly compatible with analgesics, antibiotics, etc. 
The composition of the present invention was tested in several hundred 
patients suffering from AIDS or precursors or secondary diseases related 
thereto. Hereinafter the results of these clinical tests are briefly 
summarized. 
The composition of the present invention prepared as described in Example 1 
exclusively was administered subcutaneously to several hundred patients 
suffering from tumors resulting from immune deficiency and others, such as 
those caused by HIV as an alternative therapy for the treatment of tumors. 
A favorable reaction and cessation of future growth was found in about 80% 
of the cases while in about 40% of the treated cases a complete cure was 
observed. In those cases with a positive response, the pharmaceutical 
composition of the invention did not act on the tumor but rather on the 
immune system. It can be inferred that these tumors were neoplasies due to 
immunity deficiencies because of shocks and/or stress of different origin 
depending on the patients psycho-immune-pathologies. 
In another clinical test-series 10 patients suffering from AIDS were 
treated subcutaneously with the composition prepared according to Example 
1. Six of these patients were addicts and four homosexuals or bisexuals. 
All patients showed positive ELISA, positive Western-Blot, with poor 
general physical conditions and T lymphocytes recount CD4 (cluster 
denomination four) between 130 and 200. 
The application of the composition prepared according to Example 1 in the 
treatment period of 40 days resulted in the increase of CD4 to 1300-1400 
with a remarkable improvement of the general health condition of the 
patients so that it could be assumed that the new population of T 
lymphocytes was not infected. 
Ninety days after the application of the treatment seven out of the 10 
patients showed negative Western-Blot, with normal recount of CD4 and 
excellent general conditions (e.g., a state of well being which did not 
exist when the treatment began; disappearance of prior symptoms of pain, 
discomfort, adenopathies, etc.). The seven patients appeared as if they 
never had AIDS. 
One of the parameters of AIDS diagnosis is the recount of CD4 (cluster 
denomination four). In normal health conditions the CD4 value amounts to 
1400-1500, in serious cases it is 800-400 and in very serious cases it is 
decreased to under 300. 
Another characteristic parameter of the AIDS diagnosis is the Western-Blot 
(W.B.) which is negative in healthy persons but positive in persons having 
AIDS. 
In another clinical test-series 100 patients suffering from AIDS were 
treated subcutaneously with the composition of Example I. Clinical 
recuperations wherein the CD4 increased to a normal count, and the 
Western-Blot was negative, were observed in such patients. 
The aforesaid are just of an illustrative character to show the scope and 
effectiveness of the present invention. However, it is to be understood 
that within the scope of the invention as indicated herein, the invention 
may be practiced in ways other than explicitly described hereinabove 
without departing from the intended scope of the invention. It is by no 
means intended to limit the scope of the present patent application to the 
above disclosure. 
EXAMPLE 1 
0.00324 g of phospholipase A.sub.2 obtained from the dried venom of 
Crotalus durissus terrificus and 0.0004 g of giroxina obtained from the 
same snake venom were dissolved while stirring in 20 ml of a 0.17 molar 
sodium chloride solution containing 1 ml of a 0.1% phenol solution (or 
another suitable preservative known in the art such as 0.1% THIMEROSAL) 
formed in a 0.17 molar sterilized sodium chloride solution under gentle 
stirring at a temperature of 76.degree. C. for 30 minutes; such a small 
amount of phenol would not be harmful. The solution thus obtained was 
placed in a vial of 25 ml capacity made of transparent glass, stoppered 
with a hermetic rubber cover and provided with an additional aluminum 
cover. 
EXAMPLES 2-9 
These Examples comprise the synopsis of 8 clinical histories randomly 
selected from more than 100 case histories. In Table 1, the patients 
(suffering from AIDS and testing positive for HIV) are identified by their 
initials, their sex and age, their previous symptoms, the method of 
treatment (dosage of SIIF and frequency), and the results experienced. The 
evolution of the condition of the patients is disclosed as well. 
TABLE 1 
__________________________________________________________________________ 
SICK PATIENTS WITH AIDS TREATED WITH SIIF.sup.2, WHO COMPLETED 120 DAYS 
OF TREATMENT 
DATE 
SEX AND AGE 
PREVIOUS SYMPTOMS LABORATORY 
SIIF.sup.3 
RESULT 
STATUS AS OF MARCH 
__________________________________________________________________________ 
1992 
03/87 
M 66 Fever, Depression, Asthenia, Polyadeno- 
Elisa.sup.4 + WB + 
0.30 mil. 
Elisa - 
Clinical cured. 
O.S. pathies. (Addict) C.D.4 200 
120 days 
C.D.4 800 
(More than 5 years of 
treatment) 
04/87 
M 49 Mononucleosis, Polyadenopathies. 
Elisa + WB + 
0.30 mil. 
Elisa - 
Clinically cured. 
B.G. (Homosexual and Drug addict) 
C.D.4 250 
120 days 
C.D.4 700 
(More than 5 years of 
treatment) 
05/87 
M 58 Depression, Diarrhea, Loss of Weight, 
Elisa + C.D.4 280 
0.30 mil. 
Elisa - 
Clinically cured. 
R.D. Asthenia. (Homosexual) 120 days 
C.D.4 650 
(More than 5 years of 
treatment) 
05/87 
F 18 Polyadenopathies, Depression. (Addict) 
Elisa + C.D.4 300 
0.30 mil. 
Elisa - 
Clinically cured. 
C.L. 120 days 
C.D.4 750 
(More than 5 years of 
treatment) 
06/87 
F 29 Loss of weight, Diarrhea, Fever, Depression. 
Elisa + C.D.4 290 
0.30 mil. 
Elisa - 
Clinically cured. 
A.M. (Addict) 120 days 
C.D.4 700 
(More than 5 years of 
treatment) 
07/87 
M 36 Convulsions. Asthenia, Diarrhea. Poly- 
Elisa + WB + 
0.30 mil. 
Elisa - 
Clinically cured. 
L.G. adenopathies. (Homosexual) 
C.D.4 310 
120 days 
C.D.4 800 
(More than 5 years of 
treatment) 
08/87 
F 30 Depression, Stress, Cephalalgia. Asthenia. 
Elisa + C.D.4 320 
0.30 mil. 
Elisa - 
Clinically cured. 
M.F. (Homosexual and Addict) 120 days 
C.D.4 730 
(More than 5 years of 
treatment) 
02/88 
M 31 Psychotic conduct. Polyadenopathy. Notable 
Elisa + WB + 
0.30 mil. 
Elisa - 
Clinically cured. 
C.A. Loss of Weight. (Bisexual and Addict) 
C.D.4 400 
120 days 
C.D.4 700 
(More than 5 years of 
treatment) 
__________________________________________________________________________ 
.sup.2 NON-SPECIFIC IMMUNOGENIC FACTOR STIMULATOR. 
.sup.3 NONSPECIFIC IMMUNOGENIC FACTOR STIMULATOR. 
.sup.4 ELISA = enzymelinked immunosorbent assay. 
In the first column of Table 1, the date refers to the month treatment 
began. The second column refers to the sex, age and initials of the 
patients. In the third column, the symptoms of the patients at the 
beginning of treatment are shown. The fourth column refers to laboratory 
data at the beginning of treatment. In the fifth column, the first 120 
days of treatment are referred to (the entry of "0.30 mil. 120 days" means 
0.30 ml of the composition prepared according to Example 1 administered 
daily for 120 days, "SIIF" refers to "Stimulator Immunogenic Inespecific 
Factor" (the composition of the present invention)). The sixth column 
shows laboratory results after the treatment in the fifth column. In the 
seventh column, "Clinically cured" means that CD4 counts increased to 
normal by March 1992 with repeated treatments (0.30 mil. 120 days) at 
least once a year. 
The following are Elisa results for the patients in 
TABLE 1 
______________________________________ 
Patient Elisa 
initials Date results* 
______________________________________ 
O. S. 3/5/87 positive 
O. S. 7/6/87 negative 
B. G. 4/12/87 positive 
B. G. 8/15/87 negative 
R. D. 5/20/87 positive 
R. D. 9/12/87 negative 
C. L. 6/3/87 positive 
C. L. 10/2/87 negative 
A. M. 6/10/87 positive 
A. M. 10/10/87 negative 
L. G. 7/5/87 positive 
L. G. 11/5/87 negative 
M. F. 9/5/87 positive 
M. F. 1/10/88 negative 
C. A. 2/14/88 positive 
C. A. 6/12/88 negative 
______________________________________ 
*Investigation of IgG specific antibodies against HIV (HTLVIII-LAV), 
antigens are "Env" and "Core" proteins obtained by means of recombinant 
DNA 
EXAMPLE 12-21 
These examples present 10 cases which were randomly selected from 119 case 
histories. The patients (suffering from AIDS and testing positive for HIV) 
in Table 2 are again designated by their initials, their age and sex, the 
laboratory results obtained prior to treatment (column 5) and the clinical 
aspects before treatment (column 6). The treatment control results, the 
clinical observations and the general observations are disclosed as well. 
In the column headed "Treatment" the term "0.30 ml. (daily) 60 days" means 
"0.30 ml of the composition prepared according to example 1 administered 
daily for 60 days; the numbers "0.18 ml", "0.20 ml" and "0.25 ml" are to 
be interpreted accordingly. The term "no control by patients' reasons" 
means that the patient did not agree to the implementation of necessary 
controls (e.g., CD4 counts, ELISA, and Western-blot). In the fifth column, 
ELISA followed by a number or "several" means that more than one ELISA was 
conducted. In columns 8-11, the number of days refers to the number of 
days after initiation of therapy. In columns 8-9, all "ELISA" indicated 
were positive. "W. tried classification" refers to Western-blot counts. 
3 TABLE 2 
- SUMMARY OF 10 CLINICAL HISTORIES RANDOMED FROM 119 CASES TREATED WITH 
THE 
PRODUCT PREED ACCORDING TO EXAMPLE 1 AGAINST A.I.D.S. 
W. TRIED LABORATORY CLINICAL CONTROL 
EXAMPLE CODE: AGE