JPS58148826A - Glycoprotein and remedy for tumor - Google Patents

Abstract

NEW MATERIAL:A glycoprotein having the following properties: Molecular weight: 12,000-17,000. Color reactions: Coloring of the protein by the Lowry reaction, coloring of the peptide bond and the amino acid by the ninhydrin reaction after the hydrolysis with hydrochloric acid and coloring of the saccharide by the phenolsulfurinc acid reaction. Properties and solubility: White powder. Soluble in water and aqueous solution of NaCl, etc. and slightly soluble in benzene, etc. Saccharide content: 27-33%. Isoelectric point: 4.2-7.3. Adsorbable in a 0.01M phosphroric acid buffer solution (7.2pH) by specific fractionation procedures. USE:A remedy for tumors. PROCESS:A glycoprotein (CBx) having a very powerful and selective cytotoxic action on the tumorous cells and differentiation inductive action on the recovery of the tumorous cells to the normal cells is obtained by subjecting an extract or cultivation supernatant liquid of a reticuloendothelial cell, lymphoblast, leukemic cell or fibroblastic cell of a warm-blooded animal to a salting out treatment, ion exchange chromatography, gel filtration, affinity chromatography or dialysis, etc. A remedy for tumor is obtd. using said CBx as a main constituent.

Description

[Detailed description of the invention] The present invention relates to reticuloendothelial cells and lymphoblasts of ailing animals.

Tumor therapy containing as main components a novel sl protein and glycoprotein extracted from the extract or culture supernatant of leukemia HJ cells or Ichitsumugi II 4 spores
[114ijuru.

In reality, there is no effective drug FT3 for treating tumors, and although many tumor-curing treatments have been developed by many researchers around the world, there are still new therapeutic agents and treatments in clinical practice. Combination therapy with long steel has been attempted. 111%
Therapeutic agents are broadly classified into cancer immunotherapeutic agents and cancer immunotherapeutic agents. Cancer chemistry #method is a so-called thin l1M poison, a
Because it exerts the effect of inhibiting cell proliferation in a non-discriminatory manner, it acts not only on tumor cells but also on normal fat cells, resulting in leukocyte count deficiency, infertility, hair loss, teratogenicity, and carcinogenesis. Since it exhibits a 1km action per application, the IK used must be strictly TM.
There are limits.

In addition, cancer immunotherapeutic agents do not directly suppress the growth of abscesses, but directly inhibit K1 by acting on the biological defense function.
It exerts its therapeutic action by suppressing PIi in cancer, so compared to cancer chemical and legal drugs, its expression in severe production is less, but it has a biological defense function in tumor patients. In many cases, there is no light remaining, and the therapeutic effect of MI is not necessarily as light as that of cancer chemotherapeutics.

The present inventor discovered that the reticuloendothelial system #I cells play an important role in the biological defense system of Ryoiso Inui! Believing that there was an oJ potential that could be used to treat Yuu, he spent many years researching it. Already, reticuloendothelial cells contain cytotoxic factors such as lymphotoxin and tumor necrosis factor, which can be expected to have a therapeutic effect on tumors. Granger et al.
ngeret al,, Ce1lular 1mmu
nology, 58%, 388-402 self, 1
978) and Carswell et al.
, A, et at,, t'roc, Natl. A
cad, Sci, LJ, S, A-, vol. 72, no. 9, 5666-5670 negative, 1975) reported by VC. 11.Recently, the present inventor has developed #
A mixed composition containing iE-treated lymphotoxy/and tumor necrosis factor, etc.
o-BreakingFactor, abbreviated as IJ CHF. ) can be easily decentralized, and it was announced that this CBP was effective against vJglJK-translocated experimental tumors (sequel fr110. 1981, November 2).
On the 2nd, I read the morning paper and went to bed. In the course of research on CBF by Jin, the present inventor further noted # in extracts or cultured cells of reticuloendothelial cells, lymphoblasts, leukemia cells, or i1 fibroblast cells of NntIIJ. We have discovered a glycoprotein that is different from the actual one, such as the A cell damage factor, and this shelf protein has an extremely strong and selective action on tumor cells, as well as a so-called differentiated eye poisoning action that restores tumor cells to normal. The present invention was completed by fully realizing the purpose of blocking the tumor therapeutic effect that is associated with cancer.

The sr protein of the present invention (I!')
lix, i.e. Carcino-Breake
It is called rX and abbreviated as CBX. ) to describe the physical, chemical, and biological properties of

+) Molecule 1t; α01M 17 phosphate buffer (pH
7, 2) was used as section m to perform gel P2M, and the molecule tftI
llI was determined, and its molecular weight was +2. ooo~1
7,000 Ruru.

2) Color reaction: The color reaction of CBX water solution #j was tested and the results are shown in Figure 1. In addition, the IJ- and di/hydrin reactions are described in Biochemistry Experiment Course, Sei, Protein Determination 1.
The phenol sulfur tlI method, anthrone sulfuric acid, naphthol sulfuric acid, indole 4A acid and tryptophan sulfuric acid reactions are described in Biochemistry Experiment 1111L4.
The Horn reaction was carried out according to the method described in Biochemistry Experiment Course, Vol. 3, Lipid Determination Method, 1971.

Lecture 1 As shown in the table above, CBx exhibits the coloration of tannin and carbohydrates, but not the coloration of lipids.

5) Properties/Solubility: It is a white powder and is easily mixed with water, aqueous sodium chloride solution, and phosphorus*J solution, and is easily mixed with yin, hequinane, and phosphorus.

It does not dissolve very well in foam.

4) @Content; CBxf) @Contained violet and its composition Subio (5piro, H, A,, Method
s 1o Easy-mology, B Building, 3-2
M fruit measured according to the method of 6 Sada, 1966), book I
The content of III+ bonds is 27-65%, and its sugar components are 17-20% hexose and 5-5% hexosamine.
7%, 7alic acids 5-6%.

5) Isoelectric point, when the isoelectric point of CBx was measured in isoelectric focusing electrophoresis using ampholine, it was 4-2 to 73.
It is.

6) Urenox Europeus Adsorbent in 001M phosphorus M buffer (pH 7,2) A in a minute VkJ operation using agglutenin-bound Sephadex.

7) CBx gel e in an aqueous solution of pH 1O1 pH 7.0 or pH 1tO at 4°C for 24 hours or more, and in an aqueous solution of pH 10 at 60°C for 3 hours or more.
The molecular weight and tumor cytotoxic activity are stable depending on the treatment method.

8) The +E ever-thin bag does not actually cause any hindrance to the city, but selectively gives a hindrance to the ``skinny bag''. The cytotoxicity of CBX is 1
1]'- IIa side rC main fecal quality pictured below.

The cells were cultured at 37° C. for 48 hours under aeration of oxygen gas containing 5% carbon dioxide, and the number of surviving cells that were not stained with trivan blue was counted and expressed as a 50% growth inhibition rate. immaterial 1
The rate is once the proliferation of 10'' KB cells is suppressed by 50 seconds.

? ) The method of Hozumi et al. (
Hozurni et aJ, Cancer Re
5earch vol. 40, 2919-2924 Sada, 1980
In a test according to 2010, it has an effect of inducing differentiation of tumor cells, that is, an effect of restoring tumor cells to normal state.

CBX can be clearly distinguished from CBF, which is composed of lymphotoxy/mtiy necrosis factor, etc. obtained from reticuloendothelial cells, lymphoblasts, leukemia cells, or limbal fibroblasts, or a mixture thereof, in the following points. , are clearly different substances. Lymphotoxin has a molecular weight of 7 [IL
OOO~9 [LOOO, 3a000~5111) 00°I Qo 00~2 ilo LI Q(1) 5 m,
It is known that snub, α, β and r-lymphotoxins exist (Cohen et al.
y of theLympk+okines J,
Academic)'ress, 1979).

Among these, α and β-lymphotoxins are clearly different from KCB x based on their molecular weights. Sararko, r-'Jnhot + Shin is a molecule @ 1cLOOO~2 [LOOO's $1
Gra
Ger et al., Ce1lular II]
M]+unology.

58%, 388-402 Sada, 1978), but C:
50
9, using Granger et al.'s method, I made 1 Kokakosan 1G, 0LIO~2 birthmarks 000noI.
J-nhotoxin is non-adsorbed to Ulex European agglutenin-bound Sephadex with αQ1M phosphoric acid, and is selective in cell damage Ff. There is no introduction.

As mentioned above, tumor necrosis factor selectively exhibits a damaging effect on tumor dI@, but its molecular weight is 54 ouo.
to 6,000, sugar content is 0% (Carswell,
E,^,et al,,Proc,INatl,A
cad.

Sci, USA, 72%, No. 9, 5666-5b
70 sada.

(1975) 1 fsk1 molecule 1! Igooo, sugar-containing N1- is 4Ω% (Japan!! Neighbor Newspaper: 1981, 8
25th of the month, morning edition), and the former 10% recommendation is
In that it is not a fence protein, the latter one with a sugar content of 40% is
Both differ from CBx in their sugar content and in their molecular weight.

Furthermore, these -cell removal factors'fra-Ir and Cf
3F has a molecular weight of about 300Ω (Nihon Keizai Shinkai, 1981, 228.
morning edition), which is clearly different from KCBx.

CBx is generally acquired as follows: 8. Mixed race reticuloendothelial cells, lymphoblasts, leukemia cells or marginal fibroblasts aJI! Salting out the extract of I or J@Nyojo,
It can be produced by using known methods such as ion exchange chromatography, gel chromatography, affinity chromatography, and dialysis. In addition, nude mouse, x4I
By using irradiated animals to propagate Hoso@ into Daisetsu,
You can also hold a large 1kK samurai.

Next to the CHx produced in this way, we will discuss its effectiveness, toxicity, usage, and use.

Experimental Tl KB (nasopharyngeal pharyngeal) cells, expression-1 (Wm
)-Bulle (Given by Pigeon Research Society, Dr. Shigeru Tsukagoshi), )-IEp
-2 (pharyngeal cancer), I-(El, liver cancer) cells (Flora), tFKdln-treated IJ-intestinal (Intes day ne) 407 cells, Girardet 4 rL1 scarlet (Girar).
d+ 1(eart) a@.

Chaog L+ver 4iid gun, Vero.

Monkey broom cell, MDCK (dog broom) tl&! (Flora Inc.) K, each, 10"
Tsukasa culture, P2S5 and L12+o (leukemia) cells (donated by Cancer Research Society, Dr. Shigeru Tsukagoshi) 105 mackerel were immediately taken, and the test substance was added to 10 whispers of calf serum.
@Jt! ! xwt, 37℃, 5 major carbon dioxide gas? Culture for 48 hours under oxygen gas aeration). After completing the cultivation, the number of surviving #I cells that do not stain with trypan flu was recorded under the microscope, and 50 small cells of the test substance were completely destroyed as control i100. did.

The test substances include CBx obtained in Example 2 (fabrication example), which will be described later, and CBx obtained by a known method (Granger, G, A, e
tal.

, Ce1lular Irmuu+ology, 3
Volume 8 + 588-402, 1978) @
In the case of Samurai 1, γ-lymphotoxin, and CHF, which were separated from CBx in Experiment 2 (Taizo sham) described later, mitomycin Cl was used. Also, lymphotoxin and CBF 1
The year is determined by a known method using the toxicity to mouse L cells as an index (Bloam, B, R, & Qrade, P,
)t, co-[Invitro methods +n
cell-mediated 1ttrnunit
y J^cademic Press, 197? Displayed by year. The results are shown in Table 2.

As is clear from the results shown in Table 2, CBx was able to improve C8) and co-workers' normal Ia bags without substantially impeding -
The tumor caused selective disability on one side. However, the damaging effect on each tumor cell [GILF] was different between CBx and CBF'.

This is Kn1. - Lymphotoxin and miteman/C
Both showed non-selective damaging effects on stopped cells and tumor cells.

Real MH2'')'Effects of Sarcoma 180 and Ehrlich cancer transplanted mice on Sarcoma N10dll
3 per mouse for ddY male mice with a body fi of 25 to 30 f.
X 10'1jll-a intracavity metastasis occurred, and the patient was observed on the day of survival. The CBx that was served in Example 1 (agricultural example) was 1
Five mice in the group died the next day after developing a tumor mFifJ! I was injected intravenously every day. The average survival time of the control group was 10 days.
It is shown in Table 3 as 0.

From the results in Table 5 and above, CB
It showed a clear anti-inflammatory effect.

Experimental Shock 5 Effects on survival day e of leukemic mice
4Q per game? 0'@Q? '7th leukemia L +21° or 106IllI! l mouse leukemia P2S5 4! J
I transplanted the seeds and estimated the number of days they survived. (Described later with 5 animals per row) [In 2 (manufacturing example), 1 Samurai CB x was intraperitoneally administered every day from the day after transplantation until immediately after death. The results were reported in the fourth column, with the average survival days of the control group being 100.

As a result of the above, CBx is mouse white layer disease 1.1210.
It showed a clear anti-tumor effect in both m-cancer mice and murine leukemia P388.

Soken-sho 4 Influencing substances on survival days of lung cancer mice 120
~25 # of BDF, Itooma mice were intramuscularly transferred with 2 x 104 Lewis lung cancer 1 m bags, and the survival days were determined. One group was made up of 6 animals, and CBx, which was prepared using a negative injection tube (described later), was injected intravenously every day from the day after transplantation until just before death.
00 and shown in Table 5.

From the results in Table 5 and above, CHx clearly showed an anti-inflammatory effect in Lewis MJaMAs mice.

Experimental Example 5 Effect on survival days of melanoma mice & lotus t3 DF of body 120-251, subcutaneous 10' mouse black # on the back of male mice! They were harvested for 816'ks and the number of days they survived was observed. A group of 7 animals was used in the experiment 2 (described later).
CBx, which had been given at the time of immunization (II), was injected intravenously every day from the day after inoculation until before death II. The results are expressed as the average survival days r for the #@ group.
It is shown in Table 6 as 100.

According to the results shown in Table 6, CBx clearly exhibited anti-cancer activity in mice bearing melanoma B16.

experiment? Effect of lj6 #Ih on lung metastasis 1
A 2-inch square piece of Lewis lung cancer was placed subcutaneously on the back of a group of 6 BLIF and fan mice with an IL of 20 to 501, and after 9 days, for 12 days, the samples obtained in Example 1a Example 1 (negative example) described below were placed. C
Bx and CBF were injected intravenously once a day, and 1121 days after transfer, the weight of the primary tumor and the number of lung metastatic nodules were measured by Wexra (
Wexler, H.) method (J. Natj, Ca.
ncer In5tjtute, 3441.64
1 negative, 1964) K was calculated accordingly. The results are shown in Table 7.

Table 7 The results in the table are expressed as mean value±IIA standard error.

Fist risk *There is a statistically significant difference compared to control # at 5% or less.

Compared to the control group, the Sabagan risk was less than 1 whisper (significant!
! can be.

The results above showed that CBx suppressed the primary lung cancer and all metastases very well, but CBF had little effect on lung metastasis.

Experimental Example 7 - Tumor Cell Differentiation 04 Effects (Hozumi, M, et al.
,, CancerResearch, 404! ,
2919-2924, 1980). Acute osteogenic leukemia cells (M-1 cells, provided by Dr. Motoo Hozumi, Saitama Cancer Center) SX + O''- were added with the test substance in advance, and twice the amount of amino acids and vitamins as usual. The cells were suspended in Eagle's medium 11 containing 10 Rauno's serum and cultured for 48 hours at 57° C. under a 5 minute gas aeration containing carbon dioxide. After completion of the culture.

02 Whispers, containing 1 meristerene latenox particles (Dow Chemical Company), suspended in the ground, 57℃ for 4 hours, 1 trowel, and a small grain that became able to eat verticles.
The cells were counted under the microscope and the differentiation rate was calculated from the ratio of the numbers of both images, and the results are shown in Table 8.

Table 8 CBx has a toxic effect on differentiation.

Experimental Example 8 Pyrogenicity test According to the test method described in the Japanese Pharmacopoeia, CBx, which was delivered to a white bride in a 5-carrier, was injected intravenously at a rate of 100 per animal, and 3 hours later, the rectal Thermocouple body I! The results measured using the i1 meter are shown in Table 9.

Table 9 CBx was not pyrogenic.

Experimental Example 8 Toxicity test (single administration)
#10 animals were used in Experimental Example 2 (meat production), which will be described later.
Bx (i-injected intravenously and observed the number of dead bacteria for 7 days 1
As a result, even after administering CBxICLQOO/hour, all 10 animals survived without any change in general symptoms.

9 Poison test (continuous administration for 60 days) Body weight 20-2
5JJ IJD F, 10 mice per group,
After iE implementation Reli 1 (Samurai CBx for manufacturing 30
Daily intravenous injections, number of deaths, 14 subtypes + hours, and all general symptoms were performed. Body weight was measured between 9:00 am and 10:00 am, and 129 of the general symptoms were measured using the Ardine method (5c
ience, vol. 156.

123, 1962) % in, 20.30
I did it every day and every month. M fruit and shite, CkJx, 1
．． There were no fatalities during 50 days after administration of ODD units/IF/day, and no difference was observed in body mass III circulatory toga compared with the control group. Regarding general symptoms, no abnormalities were observed in the control group and the same aircraft.

As is clear from the above experimental examples, CHx exhibits a selective growth-inhibiting effect on abscess cells (cancer 1!@) and normal 11.
It not only has the effect of 41jiK differentiation and significantly suppresses cancer metastasis, but also has an extremely wide variety of -flrK
Although they have rarely been effective, they are extremely safe and extremely useful in the treatment of tumors, even at doses much higher than those of the tKs that exhibit these pharmacological effects.

CBx is available in 1 commonly used administration methods such as injections, amends, nasal drops, temporal preparations, external preparations, oral preparations, and l1Jii.
It can be used for & administration preparations, intravaginal administration preparations, etc.

In addition, considering its safety, the daily therapeutic dose of CBx for adults is not yet determined until it is determined, but it is α5 units to 50 [LOOG], and more preferably, it is locally applicable (in α5 to aooo, 20 to too for all injections such as intravenous injection, intramuscular injection, cloO, 50 to 50 α000 units for light administration, and increase or decrease as appropriate depending on usage and symptoms. It's a monkey.

CBx may be any conventional pharmaceutical carrier, base or excipient IV3.
Medical use with cutting and customary methods! ! You can erase and zero. For light administration, there are capsules, tablets, tablets, etc., for intrarectal administration, there are 4J for rectal administration, for example, for intrarectal administration, there are water-soluble injections, and for the dosage, there are various types such as distilled water for injection. do! Use freeze drying! Isho shooting,
As an external preparation, it is preferable to use it as a soft 1t11i]I or as a lawn thin. In addition, it can also be used as eye drops], nasal drops, or inhalation powder. The implementation is shown below, but the method is not limited to this.

True flow 1 (manufacturing example) human peripheral lymphocytes 2 x 10"l 1 of 4,000d
Phytohaemaggluteni/(manufactured by Difco) was suspended in Eagle's medium containing 0.0 whisper calf serum, and 50 μg/xl of phytohemagglutenic acid was added at 57° C. under 5% charcoal gas and aerobic gas aeration at 48:00 I. 'ii1 culture. After completing the culture, remove the supernatant.
(40-8 after dialysis against NM phosphorus
0 whisper m ammonium salt precipitation 1111 minutes. This amount was dialyzed again into the above-mentioned phosphorus #IIItaI solution.
100 (Pharmacia?f) for E' (Gel FiM)
A fraction of molecules 112,000 to 17,000 in row t 6Ln was obtained and designated as the crude CBxj fraction, and the 7S: fraction eluted before that was designated as the crude CBF fraction. Crude CBx&Or,') Rex European agglutenin (Maruzen Sekiyu) adsorbed on iM Gosef7dex, rLsM fucose-containing αot
After elution with M+7 separation buffer, the fuco-st-t- was removed by dialysis, applied to agglutenin-bound Sephadex, and phosphate buffer solution (pH 7,
2) osWtt-CBx was extracted using a so-called gradient method in which Kj% was gradually reduced. In this way, CBxt
Ogg#t-obtain. The CBx activity obtained by the above summation was in IQOOO units, and the specific activity of Ii white 1mpfi was 10,000 degrees/wa1.

Implementation fi12 (Production example) Human BALL-1d@ (Miyosbi, N
ature, vol. 117, pp. 843-844, 197
7 years) 9 X 10"@i 1.800 gj of 10 calf serum containing Eagle j@ floating on the ground, Sendai virus f 9 X 10' pfu added, 67°C, 5% carbon dioxide gas containing , 48Qf'!l culture 11&, top?'1flij!JliiiM 1 Noah 1rffi[
Therefore, Fl was made. In this way, CBx720μtt
-Can serve as a samurai. The activity of CBx thus obtained is Z920 units, and the specific activity is +1.000%/nip.

Real Jll meat 6 (in production) Human fiber JJ cells 70 7000 cells (70 companies) 5
X 10" @ t-600d was suspended in Eagle J @ ground containing 10% calf serum to finalize phytohemagglutinin.
50 μy/xlK at 57°C, 5 min.
The supernatant was purified according to the method of Example 1. In this way, CBx 60μi5r can be used. The activity of CHx thus determined was 480 degrees, and the specific activity was a000 degrees/'wg.

*Example 4 (water bath injection) CBx 20 to 000 sodium chloride 9. Oj Total t i, 000j + 1 substance and chloride in distilled water for injection) After weighing and mixing lium, dissolve in 500 xi K of distilled water for injection, and then add total tt with distilled water for injection.
Let t,oood. This water bath solution was aseptically passed through a membrane filter, and the e-liquid was sterilized through glass valleys 4, each of which was filled with light for 2 d, to give a water-soluble injection.

Example 5 (Freeze-dried injection) CB x xoc4ooo 20% human serum albumin 10 II + 7 total volume with distilled water for injection (t, on-mouth dCB x H chloride) I) Press the ram and mix, then prepare for injection Distilled water 500xlK predetermined amount of human disintegrated albumin t-added water bath solution KI@dissolved, lf
! [Total 1ifr1,001 with distilled water for injection]t/. This water bath liquid was passed through an inert Kf' membrane filter and filled with light into a glass valley with 2xl ridges.
Freeze dry. V this! After 5 seconds, it was made into a lyophilized powder preparation.

Example 6 (Eye drops) CHx too, ooo Sodium monoxide 5t Globutanol 5# For injection #Total 111 mouths in distilled water 001 f: Weigh each ingredient and add to 950 ml of distilled water for injection. Dissolution - [ru. All 1 was changed to +,000d, and 1 was broken aseptically using a membrane filter to make an eye drop, 5A! Residue 7 (Suppositories CBx IOQ, 000% polyethylene glycol 1500 250 #Polyethylene glycol 4000 approx. 750 #1.000j+ Weigh each of the above components, and calculate the whole tone of CBx and polyethylene glycol 1500, and polyethylene glycol 4
0000500I We will meet each other and trust each other.

Saral! M! :l polyethylene glycol 4000i
In addition, all warehouses! t1, 0ΩOt, and a 5,000 square rectal excision was made using the bath melting method with good agreement.

Example 8 (Nasal Drop) 0 CHx 10 (LOOO4ilQ Sodium Chloride 5 to Talolophthanol 5 to Distilled Water Total ii 1,000 ILt) Each of the above components was weighed, dissolved in 950 d of water and left to cool.
Dilute to 11,000m total and use as nasal spray.

Example 9 (Enteric coated tablet) Ctlx LOOFILO00$fQ Milk II 6 hairs Potato #flour Approximately 3 nops Polyvinyl alcohol 407 Magnesium stearate 50 tons 1.000 were weighed in each of the above component warehouses, and then each of the CBx and milk rights warehouses, Mix about half of the potato starch and potato starch, and add the remaining potato flour to a volume of 94.0 g and mix uniformly. A water bath solution of polyvinyl alcohol is added to this mixture, and granules are prepared by a wet granulation method. The granules are dried and compressed with magnesium stearate to produce tablets of 20Q liter each, which are then coated with methylcellulose 7 talate. II
It was assumed to be a k-resolved fracture.

Fruit 7HM10 (ointment) CBx 10G, 000% wave paraffin 101 petrolatum / illusion 1.00Of 1.0001 The above ingredients are suppressed by n, CBx is mixed with wave paraffin, and 500 p of petrolatum is added. Mix well. Add the remaining petroleum jelly, add all*k i,
000 t, and mixed well to obtain a soft consistency.

(1 other person) Procedural Amendment L Matters Fl-No. 1. (Japanese Patent 1 [No. 289
91. Master of invention, J11M white matter tumor and Wl tumor #I treatment: i hli corrective idea 1. Relationship between special pestle and investment name Iyama M ¥ Co., Ltd. 4, 1' l Rijin 5, Supplementary 11 Anchor Y combination 19fsu 7, amendment Vi8 txt 崴Reisho recommendation 12 True 6th line -' Mayu 7 Between the lines [
In addition, the molecular weight of the fIk-component factor is 5 mm OOO according to the gel certification method, and it is unstable at pH 3 or less, and Co
It has been reported that tAJ does not adsorb to Sepharose (Mjcm
a*I, K, B, & George,
E.

G, , J, I suaauaeJogy
*Volume 125, 14i71-1aj77Ji, 19
1980), or the molecular weight is 3,000, determined by the gel filtration method, and contains almost no sugar (Matt
hews, N., at al., Brjt.

J, Caxzcer, 42nd House, 411-42.
211, 1980), but their physicochemical properties are different from those of CBz. ” is inserted.

(2) Specification 11 F stripes 12 line II line and F in the darkness of the same line 12.Furthermore, as shown in the CBxd So-NK mentioned later,
Soonmnm there is a virtue% J4 characteristic in the writing action and the enemy tumor action (
, whereas the cytotoxic effect of CBx is suitable for various tumor cells, the 2-in-7 elongation has a true nature in its effect, and the cytotoxic effect of CBx is suitable for various tumor cells. CBx and interferon are not formal O.

Also, insert the following text: CBx/d antiviral #is used as interferon.

Claims (2)

[Claims] (1) m protein with the following properties: ■ Molecular weight; 1
2,000-17. L) 0 G, ■ Color reaction; Lowry reaction shows protein coloration, ninhydrin reaction after hydrochloric acid hydrolysis shows peptide bond and amino acid coloration, phenol ii* reaction, anthrone sulfuric acid reaction, indole its It shows the coloration of li class due to the reaction, Trigto-7-amino acid reaction. ■ Properties/Solubility: White powder, chlorinated in water) 1
It is 7% soluble in water and phosphate buffer, and very soluble in benzene, hequinane and chloroform.The amount of metal is 27-55%, and its composition is 17-20% hexose. %, Hexosamine 5-7%, Sial #
5-6%, ■ Isoelectric point;
x europeus agglutjnin) In a minute-operation with combined Sephadex
Adsorbent in phosphate buffer (pH 72), ■ pH 2,0, pH 04L<t! pHtto (D
Stable in an aqueous solution at 4°C for 24 hours or more, and stable in an aqueous pH solution at 60°C for 5 hours or more; ■ Substantially does not cause damage to normal cells;
selectively damages cells; ■ Tumor @ m cell differentiation; Possesses a toxic effect. (2) A tumor therapeutic agent containing m protein having the following characteristics as a product: ■ Molecular weight; t2. ooo~17. '000゜■ Color reaction; Shows the color of proteins (by Lowry reaction), shows the color of peptide bonds and amino acids in the ninhydrin reaction of hydrochloric acid hydrochloride ps*, shows the color of peptide bonds and amino acids in the phenolumi 1m reaction, , indole (]i1#R reaction, exhibits sW4 coloration by trypto-777 sulfuric acid reaction, ■Properties/48 degradability; white powder, solidified with water)
It is soluble in IJum aqueous dispersion and phosphate buffer, but hardly soluble in benzene, hexane and chloroform. ■ Sugar content is 27-33%, and its composition is 17-20% hexose. , Hexosamine 5-7%, Sial@
5-6%, ■ 21! Electrical point; 4.2-7.5, ■ Urencus europaeus αO1M phosphorus #
Adsorptive in buffer solution (pH 7.2); ■ Stable for more than 24 hours at 4°C in an aqueous solution of pH 1O, pH 7.0 or pH 1to;
Stable for more than 3 hours at 60℃ in H7.0 water f# liquid. ■ Positive g! It has a selective effect on tumor cell differentiation, without causing substantial damage to vaginal tumors.