Note: Descriptions are shown in the official language in which they were submitted.
2168263
_ I
Pharmaceutical Composition
This invention relates to a pharmaceutical compo-
sition which contains T lymphocytes as the active substan-
ce, a process for preparing the pharmaceutical composition
and the use of same for the preparation of drugs for the
treatment of viral and bacterial infections.
Today, infectious diseases caused by bacteria or
viruses are largely treated with chemotherapeutics. Anti-
bacterial chemotherapeutics are metabolites of microorgan-
isms or semi-synthetic derivatives of same (antibiotics)
or synthetically prepared compounds (sulfonamides).
Currently, as the chemotherapeutics enabling ther-
apy of virus-induced infections by inhibiting the virus
production, nucleoside analogues inhibiting the virus-
specific enzymes (polymerases) are preferably employed.
A substantial and generally familiar drawback of
the chemotherapeutics is that` resistances may occur in
prolonged and repeated treatment. In addition, side ef-
fects are frequently reported.
So far, in spite of intense efforts, no success
has been achieved in providing chemotherapeutics which
interfere causally or symptomatically with the virus- or
retrovirus-induced events of disease with significantly
substantial success. Today, it is not possible to cure
certain virus diseases such as, for instance, the acquired
immune deficiency syndrome (AIDS), the AIDS-related com-
plex (ARC) and their pre-stages, herpes, cytomegaly virus
(CMV) infections, influenza and other virus infections or
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to exert favorable influence on the symptoms thereof
chemotherapeutically.
Currently, for example, it is almost exclusively
3'-azido-3'-deoxythymidine (AZT) which is available for
the treatment of AIDS. However, AZT is characterized by
excessively severe toxicities occurring already in the
therapeutic range [Hirsch, M.S., J.Infec.Dis. 157, 427-431
(1988)].
Therefore, it is the object of the present inven-
tion to develop a therapeutic agent which may be employed
in the treatment of viral, particularly retroviral, and
bacterial infections, without resistances or side effects
occurring in prolonged or repeated treatment and pharma-
cologically relevant doses being toxic.
Surprisingly, it has now been found that a pharma-
ceutical composition exceedingly simple to prepare and
favorable in cost, which contains T lymphocytes from mam-
mal blood, spleen or thymus gland or from autoblood as
well, is highly suitable for the treatment of viral and
bacterial infections and is rapidly effective.
The T lymphocytes contained in the composition are
recovered from m~mm~l whole blood, e.g., from rabbits,
lambs or pigs, by subjecting the m~mm~l whole blood, to
which aqueous trisodium citrate is optionally added as an
anticoagulant (0.11 moles/l; 9 parts of blood, 1 part of
citrate), to a centrifugation for about 7-8 minutes at
about 4000 rpm, and separating the leukocyte boundary
layer. By separating the leukocytes on nylon wool, a con-
centrated T cell suspension is obtained. Recovery from
autoblood is effected in analogous fashion. Preferably, to
the autoblood, there is likewise added aqueous trisodium
citrate and dextrose.
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Another possibility of recovering leukocytes from
m~mm~l whole blood or autoblood consists in passing the
whole blood, to which aqueous trisodium citrate
(0.11 moles/l; 9 parts of blood, 1 part of citrate) has
been optionally added, through a commercially available
cellulose filter, washing same with PBS, subsequently
centrifuging the obtained leukocyte/thrombocyte suspension
for 7-8 minutes at 700 rpm to separate thrombocytes and
leukocytes. Again, the separation of leukocytes is effect-
ed on nylon wool. Using this process, a T lymphocyte sus-
pension particularly high in concentration is obtained.
Similarly, of course, the leukocytes may be sepa-
rated from m~mm~l or autoblood using the well-known leuko-
pheresis process.
Alternatively, the T lymphocyte suspension of the
invention may also be recovered from m~mm~l spleen or from
m~mm~ 1 thymus gland by leaching the spleen or gland with
PBS (phosphate-buffered physiological sodium chloride
solution) optionally admixed with trisodium citrate and
dextrose, centrifuging the obtained solution for about 7-8
minutes, and separating the leukocyte layer. Again, by
separating the leukocytes on nylon wool, a concentrated
T cell suspension is obtained.
For oral application, from 20 to 50 g of the
T cell suspension is added to preferably from 100 to
300 ml of a non-denaturing liquid. In the simplest case,
water may serve as the liquid, but also juices or instant
soup find application. Use of instant soup offers the
great advantage that addition of taste modifiers or sweet-
eners for better acceptance of the drug by the patient may
be abandoned.
The preparation of an administration form of the
pharmaceutical composition of the invention with the aid
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of, e.g., instant soup is effected by preparing the in-
stant soup, cooling same to room temperature, and stirring
in the appropriate amount of T cell suspension. Here,
preferably, from 20 to 50 g of T cell suspension is dis-
solved in 250 ml of instant soup. Preferably, the T cell
suspension is stored in frozen condition (in portions) and
stirred into the liquid non-thawed. This causes the
T cells to break. When using a freshly prepared T cell
suspension to prepare the administration form of the in-
vention, the suspension must be introduced into the liquid
with vigorous stirring (mixer), in order to destroy the
T cells. A precondition for the high and rapid effective-
ness determined according to the invention is that the
T cells are taken up already in broken condition.
The T lymphocyte suspension may also be applied
rectally as a suppository or as a clyster. When adminis-
tering as a clyster, the T cell suspension preferably is
applied undiluted or diluted slightly with water for bet-
ter handling.
The administered dose depends on the recipient's
age, health and weight.
Conventionally, the daily dose is from 20 to 50 g
of T cell suspension to be given in a single application.
A modification of the pharmaceutical composition,
according to the invention, also relates to a T cell sus-
pension from autoblood prepared in analogous fashion as
the T cell suspension from mammal blood, which, however,
has to be inhaled using a sprayer. Here, however, applica-
tion must be effected using about three portions over a
day, where preferably, no more than 10 ml/day should be
administered.
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Anaphylactic shocks as observed in the administra-
tion of whole blood or whole blood preparations do not
occur in oral application of the composition according to
the invention.
The pharmaceutical composition of the invention
has valuable pharmacological characteristics. It has been
found that with bacterial or virus infections, any symp-
toms of disease were no longer present as early as after
five days of application. In serious cases, primarily with
persisting infections, the T cell suspension may be admin-
istered for up to 30 days without side effects occurring.
Repeated application of the pharmaceutical composition
according to the invention does not give rise to any re-
sistances or toxic side effects.
So far, test persons suffering from viral and
bacterial infections including those with shingles, facial
herpes, Pfeiffer infectious mononucleosis (Epstein-Barr
virus), and AIDS, have been treated using the pharmaceuti-
cal composition according to the invention prepared from
mammal whole blood.
Likewise, a dog fallen sick due to bacterial in-
fection was subjected to therapy using the agent according
to the invention.
The effect of the formulation according to the
invention in retrovirus infections was determined on AIDS
test persons using the Elisa HIV p24 antigen.
Obviously, the composition of the invention inter-
feres with retrovirus-induced diseases or symptoms thereof
without affecting the regularly working natural body func-
tions.
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`_ b
Therefore, the invention is also directed to the
use of the T lymphocyte suspension according to the inven-
tion to prepare drugs for the treatment of viral and bac-
terial infections, particularly of retroviral infections
such as AIDS and ARC (AIDS-related complex).
In the following, the invention will be illustrat-
ed in more detail in the embodiments which are not intend-
ed to be limiting.
Embodiments
Example 1
The pharmaceutical composition according to the
invention consisting of 30 g of T cell suspension from
rabbit blood in 250 ml of instant soup was repeatedly
administered to a 48 year old woman having a virus infec-
tion and (4 months later) influenza:
a) Virus Infection
- Onset of infection with signs of failing
vitality and weakness, severe headache, fever, suppos-
itories taken, recuperation without fever after
2 days;
- about 8 days later, still severe headache, return of slight fever, perspiring, shivering, dis-
ability, return to recuperation after 4 days;
- visit by physician 13 days after the first signs of infections, for now, severe weakening, slight
fever, persistent, hard cough, persistent cold, and
hence, sleeplessness and constant tiredness;
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~,
Physician's diagnosis after blood sampling: Virus
infection of an extraordinarily rare virus, with poor
liver values;
reatment: Inhalation 3 x daily, nose drops, cough drops,
6 tablets of Wobenzym N (enzyme) 3 x daily, 2
tablets of Gripp-Heel 3 x daily (influenza),
3 drops of Grippe Comp. 2 x daily;
on the 9th and 10th day: total weakness in spite of
(after physician's visit) treatment as indicated,
unbearable coughing stim-
ulus
- on the 11th day: administration of first
(after physician's visit) composition of the inven-
tion
- on the 12th day: administration of second
composition, slept through
the night for the first
time, only minor coughing
remaining, cold almost
gone;
- on the 13th day: administration of third
composition, cough and
cold completely gone, gen-
eral condition regained;
- on the 14th day: administration of fourth
composition, no longer
complaints; headache gone;
undisturbed sleep at
night;
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- on the 15th day: administration Of fifth
and last composition, no
symptoms of sickness any-
more.
b) Influenza
Four month later, severe influenza recurred which
again was subjected to therapy by ingesting the composi-
tion of the invention. After only 3 days of ingestion (1 x
daily, in the evening), alleviation ensued, and after 10
days the female patient was healthy.
Subsequent to this second ingestion, the patient's
health condition had improved in such way that so far, she
did not have any viral or bacterial infect for 1.5 years.
Example 2
A 43 year old male patient suffering from influen-
za received the composition of the invention according to
Example 1.
Mr. R.H. complained about general feebleness, limb
pain, lack of appetite and slight dizziness when changing
position. On the next day, shivering appeared as well as
febrile temperatures to axillary 38.8C.
Results:
- Good nutritional state and reduced general
condition; unhealthy appearance; skin pale, warm,
slightly sweating; sufficient blood flow in mucous
membranes; head freely movable, minor headache over
both maxillary sinuses; tongue wet, slightly furred;
rear throat wall slightly reddened.
- Heart: nothing abnormal detected
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- Lungs: bilaterally aerated, discretely in-
tensified breathing noise, at both
sides paravertebrally.
- Abdomen: nothing abnormal detected
- Extremities: freely movable
- Neurology: nothing abnormal detected.
Subsequent to administering the first composition
according to the invention, there was strong sweating the
next night. On the next morning, the patient already felt
much more comfortable. After administration of the second
composition according to the invention, there was further
improvement, and after administration of the third compo-
sition on the next day, the above mentioned symptoms had
disappeared except for a slight coughing.
Example 3
~ m;n;stration of the composition according to the
invention as a clyster.
A dog fallen ill with a bacteria-induced snow
gastritis was cured by a 14 day administration of 20 g of
T cell suspension according to the invention daily. Appli-
cation was effected enterally using a syringe.
Example 4
A~m;n;stration of the composition according to the
invention in the case of shingles.
A patient fallen ill with shingles was cured suc-
cessfully by a 10 day oral administration of the composi-
tion of the invention according to Example 1.
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Example 5
A~m;n;stration of the composition according to the
invention in the case of facial herpes.
A female patient with a large area of facial
herpes induced by the Herpes Simplex Virus was cured by a
5 day administration of the composition of the invention
according to Example 1. When ingesting once per day, the
inflammation declined as early as on the third day, and on
the fifth day the inflammatory area was completed healed.
Example 6
Administration of the composition according to the
invention to a female patient with Pfeiffer infectious
mononucleosis (Epstein-Barr virus).
The composition of the invention was administered
to a female patient having Pfeiffer infectious mononucleo-
sis once per day. As early as after the second ingestion,
the fever declined considerably, and the patient felt
better. On the third day, the patient was free of fever,
and after another three days she was able to step out of
her house again.
Example 7
A~m;n;stration of the composition according to the
invention to an AIDS patient.
The composition of the invention according to
Example 1 was administered to an AIDS patient (HIV-l) once
per day for thirty days.
While in the HIV p24 antigen test (measuring range
5-500 pg/ml) conducted prior to onset of therapy p24 anti-
Il 2168263
gen was unambiguously detected, p24 antigen could nolonger be detected after completion of the therapy. This
reveals that obviously, inhibition of virus replication
had occurred.
