UDC 615.277.3:577.134.181
`
`POSSIBLE AMPLIFICATION
`OF THE ANTINEOPLASTIC ACTION OF A FOLIC ACID ANTAGONIST
`BY METHYLCOBALAMINE ANALOGS
`
`Z. P. SOFYINA, N. V. MYASISHCHEVA, F. G. ARSENYAN, A. M. YURKEVICH
`
`Oncology Research Center of the USSR Academy of Medicine, Moscow
`
`The stimulant action of cyanocobalamine on the growth of transplanted tumors of various
`animal species (Rous sarcoma of chickens, PW-2 fibrosarcoma, sarcoma 45, and SSR [spontane-
`ous sarcoma of rats] of rats, and Guerin’s carcinoma, sarcoma 180, and lymphosarcoma of mice)
`and the attenuation of the curative effect of certain antineoplastic drugs in combined application
`with vitamin B12, noted in early studies, are caused by the active biosynthesis of its coenzymes in
`the animals’ bodies. Assessment of the functional role of methylcobalamine, one of the
`cobalamine coenzymes in the growth processes of normal and tumor cells, has drawn the greatest
`attention.
`Methylcobalamine is a coenzyme of the methionine synthetase reaction, a key link defining
`the synergy of the action of cobalamines and folic acid compounds in cell proliferation processes.
`The special importance of methylcobalamine for activation of this enzyme system has been
`noted by a study of the disrupted metabolism of cobalamines in human leukoses. The poor
`effectiveness of combined cytostatic therapy in certain forms of acute leukosis involving high
`methylcobalamine concentrations in the blood has confirmed the specificity of its action in the
`body (Myasishcheva et al., 1969). The active role of methylcobalamine in cell proliferation
`processes of hematopoietic
`in healthy animals has now been established.
`tissue
`Methylcobalamine increases the number of cells synthesizing DNA, their mitotic activity, and
`the size of the proliferative pool in the spleen of mice (Golenko et al.). A significant increase in
`the frequency of hemoblastosis development in mice has been found upon combined
`administration of methylcobalamine with endogenous blastomogens. An important point in the
`mechanism of the stimulant action of cobalamines is their inductive effect on methionine
`synthetase activity. In cultures of normal mammalian cells and human tumor cells, methionine
`synthetase activity rises noticeably with an increase in cobalamine concentration in the culture
`medium (Mangum et al.; Kamely et al.). However, various types of tumor cells differ from
`normal cells in their ability, on exposure to cobalamines, to increase the biosynthesis of
`methionine needed for rapid growth (Halpern et al.; Chello and Bertino). The salvage pathway
`with the aid of cobalamine-dependent methionine synthetase, which increases the intracellular
`pool of tetrahydrofolic acid independently of the folate reductase system, is evidently the
`principal mechanism of development of methotrexate (MTX) resistance in leukosis cells
`(Myasishcheva; Sauer and Jaenicke).
`In this connection, there is a real possibility of amplifying the antineoplastic effect of this
`metabolite by combined application
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`with cobalamine coenzyme antagonists. An understanding of the mechanism of cobalamines’
`action formed the basis for directed synthesis of methylcobalamine analogs and their testing as
`potential antineoplastic compounds.
`In chemotherapeutic experiments, we studied difluorochloromethylcobalamine and methyl-
`cobalamine chloropalladate, which had exhibited activity in vitro in suppressing bacterial cell
`growth and inhibiting DNA synthesis in human embryonic fibroblast culture (Myasishcheva et
`al., 1977).
`In developing a scheme of their combined action, we considered the basic aspects of the
`physiologic action of cobalamines in the body: monitoring of folic acid compounds’ entry into
`cells and the formation of folate coenzymes, as well as the rate of cobalamine absorption by tu-
`mor cells (Burke et al.; Tisman and Herbert; Floodh and Ullberg). Accordingly, we could count
`on the selective action of the studied compounds and the possible reduction of cobalamine-de-
`pendent enzyme activity in the body. However, it was difficult to expect a significant effect from
`their isolated application. Therefore, we thought it important to assess the antineoplastic action
`of these compounds in the context of inhibition of dihydrofolate reductase activity using MTX.
`
`Materials and Methods. The studies were conducted on mice of the C57BL, CBA, BALB/c lines and BDF1/
`C57BL × DBA(2) hybrids weighing 20–25 g, obtained from the USSR Academy of Medicine nursery. The antineo-
`plastic activity of methylcobalamine analogs was studied on transplanted leukoses L-1210 and La and on solid tu-
`mors: mammary adenocarcinoma (Ca-755), cervical uterine cancer (CUC-5), and intestinal adenocarcinoma (ACA-
`TOL). As the principal object of study, we selected solid tumors, on which it is easier to detect the stimulant effect
`of methylcobalamine than on the L-1210 and La murine leukosis model with its high proliferative pool and very
`short animal lifespans.
`Methylcobalamine (CH3Cbl) and difluorochloromethylcobalamine (CF2ClCbl) were prepared by the known
`method (Wood et al., 1968), modified in the extraction section (Tachkova et al.). Methylcobalamine chloropalladate
`(MetCbl·PdCl3) was synthesized by Ye. G. Chauser’s method. Methylcobalamine was administered intramuscularly
`at 10 mg/kg daily twice during the course of treatment 96 hours apart, and CF2ClCbl was administered daily
`subcutaneously in one 500 mg/kg dose or two 250 mg/kg doses daily for five days. The poorly soluble methyl-
`cobalamine chloropalladate was administered orally in a 2% starch suspension in a daily dose of 500 mg/kg over
`five days or twice 96 hours apart. The daily dose was administered all at once or at 250 mg twice daily. MTX from
`Lederle was used at 10 mg/kg intraabdominally at 96-hour intervals.
`Our research studied the activity of cobalamine derivatives both in combined application with MTX and with a
`quinoline derivative (NSC-170319):
`
`CH3
`
`N
`
`H2N
`
`O
`
`NH
`
`C NH
`
`NH
`
`N+ CH3 · 2Br
`
`
`
`We obtained the drug from the U.S. National Cancer Institute in accordance with a U.S.–USSR agreement on
`cooperation in the area of tumor chemotherapy. According to the description provided by the American scientists,
`the drug is a methionine synthetase inhibitor (Carter et al.). The quinoline derivative was administered intraabdomi-
`nally at 5 mg/kg daily or at 96-hour intervals, which corresponds to half the maximum tolerable dosage for the
`conditions. Treatment was begun 48 h after transplantation of the tumor. The results of the exposure were assessed
`24 h after the end of the course of treatment and at various times throughout the animals’ lives. Efficacy was
`measured by the percentage retardation of tumor growth, calculated by the conventional volume, and by the increase
`in the animals’ lifespan. In each test, control and experimental groups were created so that their numbers would
`afford statistically significant minimum calculated percentage retardations
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`of tumor growth (60%) and increased mouse lifespans (25%). In accordance with these requirements, the test groups
`consisted of 6–10 mice, and the control groups consisted of 6–10 animals, depending on the tumor strain used.
`
`Results and Discussion. Our research has revealed for the first time the stimulant effect of
`methylcobalamine on the growth of the transplanted Ca-755 and ACATOL tumors, and to a
`lesser extent on the growth of CUC-5 (Table 1). The greatest tumor growth rate under exposure
`to methylcobalamine was observed when Ca-755 was transplanted to BDF1 hybrid mice,
`compared to the growth of the same tumor in purebred C57BL mice. The reproduction of tumor
`cells was stimulated during the period of methylcobalamine administration; the greatest
`difference in tumor size in the experimental and control groups was found immediately after
`discontinuation of the drug. At later times, tumor growth in mice that had received
`methylcobalamine slowed. When ACATOL was transplanted to mice of different genders, the
`tumor growth rate on exposure to methylcobalamine differed. The drug’s stimulant action was
`considerably more marked in males (see Table 1).
`As should be expected, the isolated action of methylcobalamine analogs retarded the growth
`of transplanted Ca-755 and CUC-5 tumors to a lesser extent, and only immediately after drug
`administration (Table 2).
`Our comparative assessment revealed the greatest inhibitive activity with the use of
`methylcobalamine chloropalladate. The efficacy of Ca-755 growth retardation was more marked
`in BDF1 hybrid mice compared to C57BL mice. As we have stated, it was also in BDF1 mice that
`the stimulant action of methylcobalamine was significantly more marked. In this series of tests,
`the lifespans of BDF1 mice with mammary adenocarcinoma increased 50% when they were
`exposed to CF2ClCbl and methylcobalamine chloropalladate (see Table 2). At the same time, the
`administration of methylcobalamine derivatives produced no ACATOL growth retardation effect.
`We noted a large difference in the action of cobalamine derivatives on tumors depending on the
`application regime (see Table 2). Evidently, when a single large dose (500 mg/kg) is admin-
`istered, the drugs can dissociate, with subsequent formation of an active form that stimulates
`tumor growth.
`In accordance with our conjecture, when methylcobalamine analogs were combined with
`MTX, we observed an amplification of their action on the tumor (Ca-755, CUC-5; Table 3). The
`increased antineoplastic effect resulting from combined exposure was manifested immediately
`after the drug course and especially in the subsequent period: while the effect of MTX alone was
`absent, a fairly high percentage retardation of tumor growth remained.
`
`Table 1. Effect of Methylcobalamine on Growth of Some Transplanted Tumors
`
`Tumor
`
`Tumor Growth after Drug
`Days Drug Given
`Drug
`Administration, % of Control
`after Tumor
`Dosage,
`Transplant
`µg/kg
`1 day
`7 days
`14 days
`+74
`+21
`+23
`2nd and 6th
`10
`Ca-755 C57BL
`+180
`+65
`+10
`2nd and 6th
`10
`BDF1
`
`
`
`
`
`ACATOL:
`+20
`+23
`+31
`2nd and 6th
`10
`females
`+126
`+37
`+33
`2nd and 6th
`10
`males
`Note: Here and in Tables 2–6, the plus sign denotes stimulation of tumor growth.
`
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`Table 2. Antineoplastic Action of Methylcobalamine Analogs
`
`Tumor Growth
`Retardation,
`% of Control
`
`1 day 7 days 16 days
`
`Increase in
`Mouse
`Lifespan,
`% of Control
`
`Transplant
`after Tumor
`
`Days Drug Given
`
`µg/kg
`
`Drug Dosage,
`
`Tumor
`
`Drug
`
`Ca-755
`
`CUC-5
`ACATOL
`Ca-755
`(BDF1)
`
`CUC-5
`ACATOL
`
`Chlorodifluoro-
`methylcobalamine
`(CF2ClCbl)
`
`250+250
`250+250
`250+250
`
`2nd to 6th
`2nd to 6th
`2nd to 6th
`
`30
`43
`0
`
`Combination of
`trichloromethyl-
`cobalamine with
`MetCbl·PdCl3
`
`250+250
`500
`250+250
`500
`250+250
`
`2nd to 6th
`2nd to 6th
`2nd to 6th
`2nd to 6th
`2nd to 6th
`
`90
`13
`80
`+130
`0
`
`+8
`38
`0
`
`59
`16
`23
`+15
`0
`
`
`0
`0
`
`
`20
`0
`+18
`0
`
`54
`16
`0
`
`50
`
`10
`0
`0
`
`To understand the possible mechanisms of action of methylcobalamine analogs in the
`animals’ bodies, we performed a comparative analysis of the growth of the same tumor strains
`under isolated exposure to a methionine synthetase inhibitor (quinoline derivative) and its
`combined action with MTX. The retardation of Ca-755, CUC-5, and ACATOL growth increased
`depending on the concentration of the drug. The drug was most effective against Ca-755. When
`the dosage was increased from 5 to 15 mg/kg, retardation of tumor growth increased to 40 and
`96%, respectively. However, with increasing dosage, the drug’s toxicity also increased notice-
`ably. For example, with the L-1210 and La leukosis strains, the most optimal dosage according
`to our data was 10 mg/kg, at which the animals’ lifespans increased three- to four-fold. At lower
`dosages, the drug’s effect on mice with leukoses was substantially lower. For solid tumors, our
`studies revealed no significant increase in mouse lifespans. With combined administration of the
`drug with MTX, even at low dosages (5 mg/kg), we observed an additive effect, which was
`confirmed by the increased retardation of tumor growth (Table 4). With delayed treatment (on
`the eighth day after the tumor transplant) and daily administration of the drugs over five days (5
`mg/kg of the quinoline derivative; 2 mg/kg of MTX), the results were even more demonstrative
`(Ca-755), but with the additive effect came general toxicity (Table 5).
`An increase in tumor growth retardation and the animals’ lifespan was noted with combined
`exposure to methylcobalamine chloropalladate and the quinoline derivative (NSC-176319, Table
`6). Given the amplified action of MTX when used in combination with methylcobalamine
`analogs and a methionine synthetase inhibitor, we performed combination treatment of mice with
`Ca-755 using all three inhibitors: MTX, the quinoline derivative, and the most active cobalamine
`coenzyme analog, methylcobalamine chloropalladate (see Table 6).
`The combined use of methionine synthetase inhibitors and dihydrofolate reductase resulted in
`a significant amplification of antineoplastic action, especially long after the end of treatment.
`Under these conditions, the retardation of tumor growth was 85% two weeks after discontinu-
`ation of the drugs, while at these times in the mouse groups that received each of the studied
`compounds in isolation or in two-drug combinations, the suppression of
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`
`Table 3. Antineoplastic Action of MTX and Methylcobalamine Analogs
`
`Tumor
`
`Drug
`
`Drug Dosage, mg/kg
`
`Ca-755
`(C57BL)
`
`MTX
`MetCbl·PdCl3
`MTX+MetCbl·PdCl3
`
`CUC-5
`(CBA)
`
`MTX
`MetCbl·PdCl3
`MTX+MetCbl·PdCl3
`
`Ca-755
`(hybrids)
`
`MTX
`CF3ClCbl
`MTX+CF2ClCbl
`
`10
`250+250
`10+250+250 (given
`simultaneously)
`10
`500
`10+500 (given
`simultaneously)
`10
`500
`10+500 (CF2ClCbl,
`given 3 h before MTX)
`
`Days Drug
`Given after
`Transplant
`
`2nd and 6th
`2nd and 6th
`2nd to 6th
`
`1
`day
`75
`58
`97
`
`Tumor Growth Retardation,
`% of Control
`7
`10
`days
`days
`10
`+32
`20
`14
`75
`0
`
`5
`days
`
`
`
`
`14
`days
`
`
`
`
`Increase in
`Mouse
`Lifespan,
`% of Control
`16
`0
`0
`
`2nd and 6th
`2nd and 6th
`2nd and 6th
`
`90
`+220
`97
`
`48
`
` +100
`
`65
`
`
`
`
`
`40
`+80
`40
`
`2nd and 6th
`2nd and 6th
`2nd and 6th
`
`87
`+67
`97
`
`81
`+5
`99
`
`45
`+21
`74
`
`67
`5
`67
`
`
`
`
`
`0
`0
`40
`
`
`
`
`
`Table 4. Action of Combined MTX and NSC-176319 on Mouse Tumors
`
`Tumor
`
`Drug
`
`Drug Dosage, mg/kg
`
`Ca-755
`(BDF1)
`
`CUC-5
`(CBA)
`
`ACATOL
`(BALB/c)
`
`5
`MTX
`5
`NSC-176319
`5+5 (given simultaneously)
`MTX+NSC-176319
`10
`MTX
`10
`NSC-176319
`MTX+NSC-176319 10+10 (given simultaneously)
`MTX
`10
`NSC-176319
`5
`MTX+NSC-176319
`10+5 (MTX given 20 min
`after NSC-176319)
`
`Days Drug Given
`after Tumor
`Transplant
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`
`Tumor Growth Retardation, % of Control
`1
`5
`7–8
`10
`14–16
`day
`days
`days
`days
`days
`46
`9
`19
`19
`23
`18
`8
`41
`29
`30
`81
`62
`66
`43
`31
`69
`
`74
`
`65
`20
`
`55
`
`31
`88
`
`84
`
`75
`45
`53
`44
`
`
`12
`27
`30
`
`
`65
`43
`40
`
`
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`
`Table 5. Action of MTX and NSC-176319 Used Together on Ca-755 Growth in Mice
`
`Drug
`
`Drug Dosage, mg/kg
`
`Days Drug Given after
`Tumor Transplant
`
`MTX
`NSC-176319
`MTX+NSC-176319
`
`2
`5
`2+5 (given
`simultaneously)
`
`8 to 12
`8 to 12
`8 to 12
`
`Tumor Growth
`Retardation, % of Control
`1 day
`3 days
`+13
`12
`12
`+8
`76
`79
`
`Ratio of Number of Dead
`Animals to Number in
`Group
`1/6
`0/6
`5/6
`
`Table 6. Action of MTX, NSC-176319, and Combined Trichloromethylcobalamine with Palladium
`on Ca-755 Growth in Mice
`
`Drug
`
`Drug Dosage, mg/kg
`
`Days Drug Given after
`Tumor Transplant
`
`Tumor Growth
`Retardation, % of Control
`
`Increase in Lifespan,
`% of Control
`
`14
`0
`0
`23
`
`8
`
`0
`
`20
`
`
`
`2 days 8 days 14 days
`
`2nd and 6th
`2nd and 6th
`2nd and 6th
`2nd and 6th
`
`2nd and 6th
`
`2nd and 6th
`
`99
`37
`75
`90
`
`99
`
`99
`
`2nd and 6th
`
`100
`
`51
`7
`40
`
`58
`
`58
`
`88
`
`95
`
`0
`+29
`13
`4
`
`5
`
`44
`
`85
`
`
`
`
`
`10
`5
`250
`5+250 (given
`simultaneously)
`250+10 (MTX given 20
`min after MetCbl·PdCl3)
`5+10 (MTX given 20 min
`after NSC-176319)
`5+250+10 (NSC-176319
`and MetCbl·PdCl3 given
`simultaneously, MTX
`given 20 min afterward)
`
`
`
`MTX
`NSC-176319
`MetCbl·PdCl3
`NSC-176319+
`MetCbl·PdCl3
`MetCBl·PdCl3+MTX
`
`NSC-176319+MTX
`
`NSC-176319+
`MetCbl·PdCl3+MTX
`
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`
`tumor growth was practically nonexistent. However, we should note that toxicity increased
`simultaneously. We also found that the action of the drug combination varied substantially
`depending on the order of administration of the drugs and the intervals between them. For
`example, the simultaneous administration of NSC-176319 and MTX proved significantly less
`toxic to the body than their administration three hours apart, with equal antineoplastic effect.
`Thus, the results of the experimental research confirm our conjecture that the antineoplastic
`action of MTX can be amplified using methylcobalamine analogs and a methionine synthetase
`inhibitor. This opens up a new approach to therapeutic action on tumors using antagonists to a
`physiologic regulator of folic acid compound metabolism in the body. We have established the
`antineoplastic activity of cobalamine coenzyme antagonists. However, the activity of the studied
`methylcobalamine analogs, which inhibit certain metabolic links, is insufficient for full and
`prolonged retardation of tumor growth. The antineoplastic action of cobalamine coenzyme
`analogs can be significantly amplified by their combined use with MTX. Our experimental data
`indicate the advisability of clinical testing of the efficacy of similar combinations. Our principal
`objective at present is to develop an optimal regime of combined tumor treatment with these
`drugs based on comprehensive analysis of the mechanism of their combined action in the body.
`
`REFERENCES
`
`Golenko O. D., Myasishcheva N. V., Raushenbakh M. O. et al. Vopr. med. khim., 1974, No. 5, pp. 549–554.
`
`Myasishcheva N. V. “Characteristics of the Metabolism of B12 Compounds (Cobalamines) in Leukoses.” Doctoral
`dissertation, Moscow, 1972.
`
`Myasishcheva N. V., Levina, G. D., Lorne, Yu. I., et al. Probl. gematol., 1969, No. 4, pp. 20–25.
`
`Myasishcheva N. V., Golenko O. D., Kuznetsova, L. Ye., et al. Vopr. med. khim., 1977, No. 5, pp. 622–628.
`
`Tachkova Ye. M., Rudakova, I. P., Myasishcheva N. V., et al. Bioorg. khim., 1976, No. 4, pp. 535–541.
`
`Burke G. T., Mangum J. H., Brodie J. D. Biochemistry (Wash.), 1971, v. 10, p. 3079–3085.
`
`Chello P. I., Bertino J. R. Biochem. Pharmacol., 1975, v. 25, pp. 889–892.
`
`Floodh H., Ullberg S. Int. J. Cancer, 1968, v. 3, pp. 694–699.
`
`Halpern B. C., Clark B. R., Hardy D. N., et al. Proc. Nat. Acad. Sci. USA, 1974, v. 71, pp. 1133–1136.
`
`Kamely D., Littlefield J. W., Erbe R. W. Ibid., 1973, v. 70, pp. 2585–2589.
`
`Mangum J. H., Byron K., Murray J., et al. Biochemistry (Wash.), 1969, v. 8, pp. 3496–3499.
`
`Sauer H., Jaenicke L. Blut, 1974, Bd 28, S. 321–327.
`
`Tisman G., Herbert V. Blood, 1973, v. 41, pp. 465–469.
`
`Wood Y. N., Kennedy T. S., Wolfe R. S. Biochemistry (Wash.), 1968, v. 7, pp. 1707–1713.
`
`POSSIBILITY OF POTENTIATING THE ANTINEOPLASTIC ACTION OF FOLIC ACID
`ANTAGONIST BY METHYLCOBALAMINE ANALOGUES
`Z. P. Sofyina, n. V. Myasischeva. F. G. Arsenyan, A. M. Yurkevich
`
`Summary. The effect of methylcobalamine and its analogues (difluoro-chloromethylcobalamine — CF2ClCbl
`and methylcobalamine chloropalladate — MetCbl·DdCl3) on the growth of transplantable tumours in mice;
`mammary adenocarcinoma (Ca-755), carcinoma of the uterine cervix (CUC-5), carcinoma of the intestine
`(ACATOL) was studied. The activity of the cobalamine coenzyme analogues was investigated when used alone or
`combined with inhibitors of dehydrofolate reductase and methyonine synthetase. The results of the experiments
`indicate a stimulating effect of methylcobalamine on the growth of transplantable solid tumours in the animal
`organism. The antitumour activity of the methylcobalamine analogues studied was found to be higher in combined
`application with methotrexate. The most effective inhibition of tumour growth and the longer survival of the animals
`were achieved in combined application of methylcobalamine with methotrexate and methyonine synthetase inhibitor,
`depending upon the scheme of administration.
`
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`
`CONTENTS
`
`XL Session of the General Meeting of the USSR
`Academy of Medicine, April 4–7, 1978, Moscow
`(materials to be published in Nos. 1 and 2, 1979) ............ 3
`
`
`
`Sidorenko, G. I. On the Work of the Presidium of
`the USSR Academy of Medicine in 1976 and 1977 ......... 3
`
`Kosyakov, P. N. Immunology in Contemporary
`Medicine......................................................................... 14
`Baroyan, O. V., and Kaulen, D. R. Current Views
`on Developmental Paths of Immunology (Problems
`and Prospects) ................................................................ 21
`Solovyov, V. D. Some Aspects of Antiviral
`Immunity ........................................................................ 30
`Ado, A. D. Current Problems of Allergic Reactions ...... 34
`Lopukhin, Yu. M. Primary Immunodeficiencies and
`Methods of Correcting Them ......................................... 43
`Petrov, R. V. Theoretical Foundations, Status, and
`Prospects of Clinical Immunology ................................. 65
`Discussion of the Summary Report of the Presid-
`ium of the USSR Academy of Medicine for 1976–
`1977 at the XL Session of the USSR Academy of
`Medicine Held April 4–7, 1978 ...................................... 68
`
`Sofyina, Z. P., Myasishcheva, N. V., Arsenyan,
`F. G., and Yurkevich, A. M. Possible Amplification
`of the Antineoplastic Action of a Folic Acid
`Antagonist by Methylcobalamine Analogs .................... 72
`
`CONTENTS
`
`The XL Session of the General Meeting of the
`Academy of Medical Sciences of the USSR Held in
`Moscow on April 4—7, 1978 (The materials will be
`published in Nos. 1 and 2, 1979)
`Sidorenko, G. I.: On the Work of the Presidium of
`the Academy of Medical Sciences of the USSR in
`1976—1977
`Kosyakov, P. N.: Immunology in Current Medicine
`
`Baroyan, O. V., Kaulen, D. R.: Current Views on
`the Ways of Immunology Development (Problems
`and Prospects)
`Solovyov, V. D.: Aspects of Antiviral Immunity
`
`Ado, A. D.: Current Problems of Allergic Reactions
`Lopukhin, Yu. M.: Primary Immunodeficiencies
`and Methods of Their Correction
`Petrov, R. V.: Theoretical Foundations, Present
`State and Prospects of Clinical Immunology
`Discussion on the Summary Report of the Presid-
`ium of the Academy of Medical Sciences of the
`USSR for 1976—1977 at the XI Session of the
`Academy of Medical Sciences of the USSR Held
`on April 4—7, 1978
`Sofyina, Z. P., Myasishcheva, N. V., Arsenyan, F.
`G., Yurkevich, A. M.: Possibility of Potentiating
`the Antineoplastic Action of Folic Acid Antagonist
`by Methylcobalamine Analogues
`
`ABSTRACTS OF ARTICLES PUBLISHED IN THIS ISSUE
`
`UDC 61:012.017.1
`“Immunology in Contemporary Medicine.” Kosyakov, P. N. Vestn. Akad. Med. Nauk SSSR, 1979, No. 1, p. 14.
`The author notes the importance of immunology to many divisions of contemporary medicine, primarily the
`immunology of infections, as well as many divisions of noninfectious immunology. Thanks to their unique
`specificity and high sensitivity, immunological methods have found wide application in various fields of biology
`and medicine. The author points out that immune reactions, which are defensive in nature, can be perverted by
`various factors and directed not only at foreign antibodies, but also at some self, normal, unaltered cell and tissue
`antigens, resulting in true autoimmune diseases.
`Bibliography: 15 citations.
`
`UDC 612.017.1:001.8
`“Current Views on Developmental Paths of Immunology (Problems and Prospects).” Baroyan, O. V., and
`Kaulen, D. R. Vestn. Akad. Med. Nauk SSSR, 1979, No. 1, p. 21.
`The authors present the basic problems facing immunology. They discuss the main problems, the possibilities of
`finding ways of purposefully regulating the body’s immune response. The authors see this possibility in the
`development of methods that regulate cell cooperation, modifying cellular microcirculation, and using antibody
`fragments. They draw attention to the value of soluble mediators of cellular immunity, lymphokines. They draw
`special attention…
`
`Lilly Ex. 2041 pg. 8
`Sandoz v. Lilly IPR2016-00318
`
`

`
`Morr:iingside TRANSLATOR CERTIFICATION
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`
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`Date: September 12, 2016
`
`To whom it may concern:
`
`I, Paul B. Gallagher, a translator fluent in the Russian and English languages, on behalf of
`Morningside Translations, do solemnly and sincerely declare that the following is, to the best of
`my knowledge and belief, a true and correct translation of the document(s) listed below in a
`form that best reflects the intention and meaning of the original text.
`
`The documents are designated as:
`• Sandoz Inc. - Exhibit 1002-00774- Exhibit 1002-00781
`
`I hereby declare that all of the statements made herein of my own knowledge are true and that
`all statements made on information and belief are believed to be true; and further that these
`statements were made with knowledge that willful false statements and the like so made are
`punishable by fine or imprisonment, or both, under Section 1001 of Title 18 of the United
`States Code.
`
`/~~
`"" Signature of Paul B. Gallagher
`
`Accurate Translation Services 24/7
`
`Lilly Ex. 2041 pg. 9
`Sandoz v. Lilly IPR2016-00318
`
`

`
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`72
`
`Sandoz Inc.
`Exhibit 1002-00774
`
`Lilly Ex. 2041 pg. 10
`Sandoz v. Lilly IPR2016-00318
`
`Lilly Ex. 2041 pg. 10
`Sandoz v. Lilly IPR2016-00318
`
`

`
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