JPH04198134A - Treating agent and preventing agent for senile dementia - Google Patents

Abstract

PURPOSE:To obtain a treating agent and a preventing agent for senile dementia, increasing choline acetyltransferase(ChAT) activity, comprising GM-CSF as an active ingredient. CONSTITUTION:GM-CSF(granulocyte macrophage colony stimulation factor) is administered alone or IL-3(interleukin-3) is administered and then GM-CSF is administered to prevent or treat various kinds of dementias based on nutrition activating action on central choline agonist neuron and ChAT activating action. GM-CSF shows nerve protrusion extending effects on nerve cells in cooperation with IL-3 or on nerve cells treated by IL-3. The dose of the compound is 0.01-300mg, preferably 0.1-100mg per adult daily.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to GM-CSF or GM-CSF and IL
-3 as an active ingredient. For more details, please refer to G.
The present invention relates to a treatment/prevention agent for senile dementia, which contains M-CSF as an active ingredient or by administering GM-CSF after administration of IL-3.

(Background to the Invention and Prior Art) With the rapid increase in the aging population, there is a strong desire to establish a treatment method for senile dementia such as Alzheimer's type senile dementia. However, although various attempts have been made to treat senile dementia with drugs, there are currently no fundamentally effective drugs. 'Alzheimer's type senile dementia is accompanied by a decline in the function of cholinergic neurons in the brain, and attempts have been made to develop acetylcholine precursors and acetylcholinesterase inhibitors, but the development of a definitive treatment has not yet been successful. . On the other hand, drugs that activate the function of cholinergic neurons and maintain their survival are thought to be effective in treating these diseases, and are being developed. Therefore, the present inventors have been conducting intensive research for many years on compounds that have a choline acetyltransferase activating effect.

[Structure and effects of the invention]

As a result, we surprisingly found that GM-CSF (granulocyte/macrophage colony stimulating factor) alone or after the action of IL-3 (interleukin-3).

It has been found that the action of GM-CSF on choline acetyltransferase increases the activity of choline acetyltransferase.

Human GM-C5F is a glycoprotein (molecular weight 22,000) consisting of 144 amino acids, and mouse GM-C5F is
It has 54% homology with SF. IL-3 is a glycoprotein with 152 (human) to 166 (mouse) amino acids! (molecular weight 25,000). GM-CSF
Both IL-3 and IL-3 are lymphokines produced by helper T cells upon antigen stimulation, etc., and have been found to act on the differentiation and proliferation of bone marrow cells.

In the present invention, GM-CSF is a mouse product produced using genetic engineering techniques, and IL-3 is a human product produced using genetic engineering techniques, but they may also be extracted from nature. The species difference does not matter. That is, it may be human or mouse, but human origin is preferred.

Next, an experimental example will be shown to describe the effects of the present invention.

i) Experimental method Nerve cells (1 x 105 cells/-) in the septal area of the mouse brain on day 15 of embryonic development were cultured in complete serum-free synthetic media (Eagle medium and Ham's
Glucose 2 was added to a 1:1 mixture of FI□ medium.
．． 25 g/l, 15mM Hepes buffer (pH 7,
4), 30nM sodium selenate, 20mM penicillin, streptomycin, 100μg/ld transferrin, 25μg/i insulin, 20nM progesterone, 20nM hydrocortisol 7-21-! J phosphate, 10μML-carnitine, 30n3.3', 5-iodide-L-thyronine, 7μg/d, tocopherol,
7μg/relative retinol, 1μM thioctic acid and 1μβ
/contains a mineral mixture) (5% C
O2, 37”C).

First, 10 units/dGM-CSF or 50 units/1dl
Culture was started with or without addition of L-3, and after 20 hours, the cultured cells were washed once, and then IO units/dGM
- CSF or 50 units/dlL-3 was added. At 0.20, 40.50 hours after the start of culture, the number of neurons having neurites was counted under a phase contrast microscope and compared with a control (no addition). The results are shown in Table 1.

Next, neurons (6×10
Sow 5 pieces/d), 1. 5. 10.50°100 units/d(7) GM-CSF was added. Addition 2 or 3
After 5 days, the medium was replaced with a medium containing the same amount of CM-CSF, and after 5 days, ChAT activity was measured according to the method of Fonnum and compared with the control group. The results are shown in Table 2.

Furthermore, in order to examine the interaction with rL-3, neurons (6×10 5 cells/relative) were seeded in the above-mentioned medium.

For the first three days, the cells were cultured with the addition of 50 units/dlL-3, and for the next two days, they were cultured with the addition of 10 units/dGM-CSF, and ChAT activity was measured.

At that time, those cultured with only IL-3 added, GM-
Comparisons were made with those cultured with the addition of only CSF and those cultured with no additive for 3 days, followed by the addition of GM-CSF. The results are shown in Table 3.

surface, mouse GM-CSF' used in this experiment, human IL
-3 is Genzyme Inc. (Boston,
(Massachusetts) was used.

1) F. Fonnui: J. Neuroch
Em, 24.407-409 (1975) ii) Experimental Results 50 units/1111L-3 was applied for the first 20 hours of the four lobes of the flexural process.

When activated, the number of cells with neurites significantly increased. Neurite outgrowth effects were observed with IL-3 alone, but not with GM-CSF alone. That is, GM
- CSF in collaboration with IL-3 or IL-3
It was found that the neurite outgrowth effect was observed in the neurons treated with the drug.

Table 2 shows the ChAT activity when various concentrations of GM-CSF were applied. It was found that ChAT activity increased as the concentration increased, but ChAT activity tended to gradually decrease at higher concentrations.Table 3 shows that when GM-CSF and I
The interaction of L-3 on ChAT activity is shown.

It was found that when GM-CSF was allowed to act after first acting on IL-3, its activating effect on ChAT activity was further increased.

The above pharmacological experiments revealed that GM-CSF has a trophic effect on central cholinergic neurons and increases ChAT activity. In addition, GM-CSF acts in cooperation with IL-3 to promote protrusion elongation and promote Ch.
It was also found that AT activity was further increased than when used alone.

Senile dementia, especially Alzheimer's type senile dementia,
There is a loss of cholinergic neurons, which has been linked to some of the symptoms of dementia. Therefore, administering GM-CSF after administering IL-3, or
Administering alone is effective for various dementias based on the nutritional activation effect on central cholinergic neurons and choline acetyltransferase activation effect.

GM-CSF has been tried to be administered to agranulocytosis patients, but no particular side effects have been found.

IL-3 has also been tried to be administered to patients with terminal cancer whose bone marrow has been irradiated with X-rays and to AIDS patients, but no particular side effects have been found.

The compounds of the present invention are effective for all diseases for which choline acetyltransferase activation is effective.

Typical diseases include various senile dementias, especially Alzheimer's disease, Alzheimer's type senile dementia, Parkinson's disease, stroke (cerebral hemorrhage, cerebral infarction), cerebral arteriosclerosis, brain disorders associated with head trauma, and sequelae of encephalitis. It is effective for the treatment, prevention, remission, and improvement of cerebral palsy, Down's syndrome, reduced attention span due to age, language disorders, decreased motivation, emotional disorders, memory disorders, hallucination-delusional states, behavioral abnormalities, etc.

GM-CSF was recently found to be produced by astroglia. It is also known that vascular endothelial cells also produce GM-CSF, and it is highly likely that vascular endothelial cells in the brain also produce GM-CSF.

In addition, it has recently been revealed that IL-3 has a trophic effect on central cholinergic neurons.
-3 has also been shown to be synthesized in the brain. Considering these facts and the above experimental results, CM-
It is highly likely that CSF alone or in combination with IL-3 and GM-CSF acts as a trophic factor for central cholinergic neurons.

When the compound of the present invention is used as a pharmaceutical for these, GM-CSF, IL-
3 is formulated to be effective in the treatment and prevention of this disease, and the active ingredients GM-CSF or GM-CSF and I
Administer L-3.

The dosage of GM-CSF or GM-CSF and IL-3 depends on the severity of symptoms; age, sex, body weight, and sensitivity differences of the patient; administration method; administration time, interval, nature of pharmaceutical preparation, preparation, Type: Varies depending on the type of active ingredient, etc., and is not particularly limited, but usually about 0.01 to 300 per day for adults
mg, preferably about 0.1 to 11001I1, and is usually administered in 1 to 4 divided doses a day.

In order to formulate a compound of the present invention, it is prepared into a dosage form such as an injection, a herbal medicine, a sublingual tablet, a tablet, or a capsule by a conventional method in the field of pharmaceutical preparation.

When preparing injections, add p)I adjusters, buffers, suspending agents, solubilizing agents, stabilizing agents, isotonic agents, preservatives, etc. to the herbal medicine as necessary, and prepare by conventional methods. Administer as intravenous, subcutaneous, or intramuscular injection. At that time, if necessary, it is also possible to freeze-dry it by a conventional method.

Examples of suspending agents include methyl cellulose, polysorbate 80, hydroxyethyl cellulose,
Examples include gum arabic, powdered tragacanth, sodium carboxymethyl cellulose, and polyoxyethylene sorbitan monolaurate.

Examples of the solubilizing agent include polyoxyethylene hydrogenated castor oil, polysorbate 80, nicotinic acid amide, polyoxyethylene sorbitan monolaurate, magrogol, and castor oil fatty acid ethyl ester.

Examples of stabilizers include sodium sulfite, sodium metasulfite, and ether; examples of preservatives include methyl p-oxybenzoate, ethyl p-oxybenzoate, sorbic acid, phenol, cresol, and chlorocresol. .

Claims (7)

[Claims] (1) A therapeutic/preventive agent for senile dementia containing GM-CSF as an active ingredient. (2) A therapeutic/preventive agent for diseases containing GM-CSF as an active ingredient and effective in activating choline acetyltransferase. (3) A choline acetyltransferase activator containing GM-CSF as an active ingredient. (4) An agent for treating and preventing senile dementia by administering GM-CSF after administering IL-3. (5) A therapeutic/preventive agent for diseases in which the choline acetyltransferase activation effect is effective by administering GM-CSF after administering IL-3. (6) Choline acetyltransferase activator by administering GM-CSF after administering IL-3. (7) The agent for treating and preventing senile dementia according to claims 1 and 4, wherein the senile dementia is Alzheimer's disease or Alzheimer's type senile dementia.