Bior& & Phorttraror/wr (1991) 45.49-54 Q Elsevier. Paris
49
Editorial
estatin, an aminopeptidase with a multi-pharmacological
inhibitor function
G Math6 lnslitur du Cattccr et d’/tttatuttoji~nnPtique 14-16 avenue Paul-Vaillanr-Couturier. 94800 Villejuif,
France
Summary - Bestatin, the dipeptide N-1(2& 3R)-3-amino-2-hydroxy-4-phenyl butanyl) L-leucine is a leucine aminopeptidase and aminopcptidase-B inhibitor. It exerts a direct stimulating effect on lymphocytes (and monocytes) viu its fixation on cell surface leucinc-aminopeptidase. and an indirect effect on monocytes (and lymphocytes) via aminopeptidase B inhibition of tuftsin catabolism. Thus it is an immuno-modifier as shown by the Japanese and also our groups: a) immunomodifier, especially stimulator, in normal young mice; b) immunorestorator and spontaneous tumor preventive agent in aged mice; c) immunorestorator in the elderly and in cancer patients and HIV infected subjects. It stimulates granulocytopoiesis and thrombocytopoicsis irt vim. and can reslore them in viva in myelo-hypoplastic man. The anti-aminopeptidase action of Bestatin also protects cnkephalins againt their catabolism, which encouraged us. with good preliminary results, to study its analgesic action and search for a preventive effect on “desaddicted” toxicomaniac relapses. Bestatin / anti-aminopeptidase
I pharmacological
action
KCsume - La Bestatine, un inhibiteur d’aminopeptidase avec un r6le multi-pharmacologique. Bestutine. lc dipepride N-[(2S, 3R)-3-rttttitlo-2-hydro.vy-4-phcnyl huronoyl) L-leucine est rut inhibireur de la leucine atninopepridase ef de l’antinopepridasc-B. II esis~c 1~1 cjjerl srimulanr direct sur lcs lyttlpltocytes (et les ntonocytes) par i’interttt~diaire de su jkarion sur lu Ieucitte attrittoltc~l)rirktsc de IN surface cellulaire. et utt effer indirect sur lcs ntonocy~cs (er les Iytttphocytes) par I’inrerttrklittire de I’initibiriott du ccmholisnre de la tufrsinc pm I’aminopcp~idase B. La Bestarine esr un ittuttut~o-tnodulant que sitt~uli~retnenr des groupes jopotwis et le tldrrc on1 tttotrtrL’ se cotttplircr enwe aulre cotnme : a) un inununotnodificateur, un srimulateur, cite: dcs souris jeunes et normales: h) un iomntltnorestualtralettr et parall&wrcnt un agent prfventif de la sur\*entte des tutncttrs spotlrutties cite,- lcs souris dgPes: c) rtn inunut~orestaurateur chez des ituntains tigt%, cites des patients rtrk+trs dc cwtcers CI cite: des sujers ittfech par le HIV. II stintrtle la ~mttulocytopoiPse et la rrombocytopoi2se in vitro CI peur IN res~aurer in vivo clre-_ des myE’lo-kypoplnsries imttraines. L’action anti-aminopepridase de la Besturine pro&e ttrtssi lcs ettk~pltalittes comre leurs cataholistncs, ce qtri nous a conduit, avec des rhtl~als pr~linrinaires encotrra,~eatt~s. d itudier /cur urtiott ~muI~~Csiqrte et Li recherciter leurs Cvenruelles capaciks de pre’renir les rechures dc toxicotnanes prhIcrbientcnr soutttis ti uttc cure de dhintoxication. Bestatine / anti-aminopeptidase
inhibiting action gives a only a potential but a very likely chance of curing several, sometimes unexpected and physiologically very different actions. Possessing
pharmaceutic
an
enzyme
/ action pharmacologique
agent,
A peptidase inhibitor
not
Bestatin is a peptidase inhibitor, a small molecular weight dipeptide, N-[(2S, 3R)-3-amino-2-hy-
50
G Mathi:
droxy-4~phenyi butanoyl] L-leucine (fig 1) which was extracted from Sr~q~oq~~s olivoreti [ 171. It was discovered by Umezawa [ 171 who also discovered that it is an inhibitor of leucine aminopeptidase {which is known to bind at the cell surface [9]) and aminopeptidase B (fig 2). The first inhibiting action (or one of the) is the direct immuno-active effect lymphocytes. The second inhibiting action, on aminopeptidase B, works by inhibiting the catabolism of tuftsin [6] (which results from immunoglobulin under the effect of carboxypeptidase [6]), tuftsin which is itself an immuno-(mainly monocytic-macrophage) stimulator.
*--.
STIMULATORY
EFFECT
Fig 1. Chemical s~ruc~urc of Bestatin. (k)-N-[(?S. 3R)-3amino-7-bydrory-4-phenylbutanoyi] t&wine (MW: 308.38).
PEPTWASE tNHlBiTORS BESTATIN AND PEPSTATIN FROM lg
BESTATIN
TUFTSIN H-Thr-Pfe-Arg-Lys-OH
=
INHIBITS TUFI’SIN
\ PEPSlATIN INHWTS \
INHIBITOR Lys-Pro-Arg \
b lo,,g* loopg”
MOUSE INHIBITORY EFFECT r->
-L&INE AMINOPEPTIDASE
Fig 2. Bestatin is: a)aminopcptidase B inhibitor. b) as ppstatin. a leucinc aminopcptidase inhibitor (1). and c) a tuftsin catabolism inhibitor.
An i~~~~o-~od~iato~ statin
and r~sto~ator Be-
it appeared that Bestatin was able to globally stimulate the normal immune functions in young mice (fig 3) [S, 81. More interestingly, we observed that it was able to restore these functions on age ~mmunodepressed mice (fig 3) 141. It not only stimulates in normal mice, T-lymphocyte functions, especially CD4+ interleukin 2
Fig 3. a. Action of Bestatin in the main immunologic functions in young mice. b. Effect of ageing on main immunologic functions. c. Effect of Bestatin according to dose on the immunologic functions in old mice.
Bestatin. an am~no~ptidas~ inhibitor
5
3
i
51
DAY AFTER TREATMENT Fig 4. Effect of oral administrationin mice of Bestatin on 3 conseculivc days on IL-I and IL-2 production by macrophages and spleen cells respectively. 1L-1 and IL-2 pr~uctions, induced irt vitro respectively by LPS and Con A. are amplified 3 and 5 days after a 3-day treatment with Beststin,
1
3
Fig 6. Effects of phospnulipase At (PLAZ) injection on the chemoluminesccnce response of peritoneal cells from immunostimulated mice. Groups of 10 mice were given, on day-a with Bestatin (5 p&mouse ip) and with or without ip injection of 0.7 units PLA?, mouse 30 min before cell harvesl. Each peritoneal cell suspension was tested in~viduatiy. The results are expressed as mV/min f SD.
5
DAY AFTER TR~TMENT Fig 5, Respective effects of Bestatin and amastatin oral administratians in mice for 3 consecutive days on peritonerl macrophage chemolumincsccnce triggered by opsonised zymosan. Fig 7. Pe~toneal cell c~moluminescence after (100 pg) tuftsin and or (25 pg) iv administration.
production (fig 4), but also B cell-antibody production (fig 3a) [5J. It can activate the mono~ytic-ma~rophagic system, increasing interleukin 1 production [fig 4) and peritoneal macrophage chemoluminescence induced zymosan (more intensely than amastatin) (fig 5), and phospholipase A2 (fig 6) or activated by tuftsin (fig 7) 13, 101. In man, it has increased, in cancer patients and NIV infected subjects, the CD&/CDS+ ratio (fig 8) 1121.
Bestatin
More interestingly, Bestatin is able to increase the CD4+ absolute number in the elderly, if the CD8+ cell absolute number is normal (fig 9) [ 101. It can aiso decrease at a certain dose (15 mgld) the number of CD8+CR3~lymph~ytes considered as suppressor cells, while it may increase them at a higher dose (30 mgld) {fig 9) [lo].
G Math6
52
Curative and preventive effects on tumors Bestatin has been shown to exert a regressive effect on murine grafted tumors f I]. It also works in combination with zinc gluconate [ll] as laro semu adjuvant therapy, but only when applied both as stricto-sensu adjuvant and neo-adjuvant treatment to surgery (before and after) (table I) [2]. In addition, when applied between the 16th and the 28th month, it prevents, especially at a high dose, the appearance of spontaneous induced tumors related to ageing, while, as seen above, it restores immunity and delays mortality (table II) [41. In man, clinical controlled trials have been conducted, of which the most interesting has concerned acute myeloid leukemia: the patients being stratified according to age [ 13, 141. There was no increase by Bestatin active immunotherapy of the ABSOLUTE NUMBERS
ABSOLUTE NUMBERS
1800 t
-..._._._._. ,___,_._.__.... ..1.”
(CD4 + , N) (CD8+,
CDB+ NKHi t
NKHt (NKH1 t N)_
Pii: fp. Effect of Bestatin according to dose on immunologic parameters, normality or decrensc.
Table 1. Proportion OF cured animals at day
.._
ml+
CD8t
N)
.
60 after surpy
especially on CD4+lymphocytes according to CDS+ cells
alone or
followed and/or preceded by Bestatin and zinc
iinnlullotherapy, I -0HP cfleinothcrapy or D-Trp-bLH-RH ho~onothe~py. -. Trca~fmW ahe Surgery Zinc gluconate + Bestatin
ffcoadjwmt
Adjutant
Neoadjrrvarrt + Adjtrvattt
0 0
0
6
21
I-OHP(l9)
40
62
66
43
D-Trp-6-LH-RH (20)
10
55
40
50
Bestatin. an aminopeptidase
inhibitor
53
Table II. Prevention by Bestatin of the incidence of spontaneous tumors appearing in FI (C57Bu x BALB/c) micebetween the 16th and 28th month of age. Untreated qed group
Malignanl lymphoma Hepatoma Leukemic lymphoma
Histological type of the tumors No of tumor bearing animals* Total No of animals per group *P
tO0 ~18bestatin-treated aged group
6 1 1
Lymphoma
$=36%
1
+)=5%
= 0.01
.
I
rate which included a rather high proportion of patients younger than 40 (and even 50). The survival rate among controls over the age of 40 is, on the contrary, low, and is elevated by Bestatin at the level of that of patients under 40 years of age [ 13. 141.
Fig 10. Synergy between rhGM-CSF and Bestatin in hemalon cultures.
survival
A pan-hematopoietic storator
stimulator
11
0 6000
and re-
The increase of in viva mice granulocyte number following chemotherapy and radiotherapy has been noted by Talmadge [IS]. We have discovered in vitro, on human CFUs, that Bestatin potentiates the effect of CFS (fig 10) [lo], and we have registered irt vivo in man, this restoration effect on granulocytes and platelets (fig 11) [lo].
150
0
Fig 11. Effects of Bestatin (applied alone to a patienl with subacute marrow aplasia after a long application of cytostatic chemotherapy).
G Math6
54
‘Enkeghalinase’
BESTATIN
lntraneuronal enkephaline
TH~OFW-WJ
>
Aminopeptidases
Release
Fig 12. Bestarin inhibits endogenous enkephaline catabolism via aminopeptidase inhibition.
Other effects, especially psychic
7 Herman ZS, Stachura Z, Opiela L. Ziemion IZ, Na-
that enkephalins are pentapeptides of the general formula tyrosine-glycine-glycinephenylalanine, the last amino acid being either methionine or leucine. Bestatin inhibits at least three aminopeptidases in the brain (fig 12) 116) and tuftsin is known to exert an analgesic effect [7]; we have searched for: a) an analgesic effect: b) a relapse inhibitory effect in toxicomaniacs after desaddiction. Our studies, which are in phase II progress, are very encouraging. It is known
References Abe F. Shibuya K, Uchida M, Takahashi K, Horinishi H, Matsuda A, lshizuka M, Takeuchi T, Umezawa H (1984) Effect of Bestatin on syngeneic tumors in mice. Gunn 75, 89 Bourut C, Chenu E, Math6 G (1988) Management of Minimum
Residlral
Malignancy
in Man.
Oxford, p 59 Bravo-Cuellar A. Homo-Delarche
Perga-
mon,
bouys S (1990)
Phospholipase
AZ,
F, Orbach-Aran in vivo im-
munomodulator. Prost Leltko Essent Fatty Acids 40, 31
8 Ishizuka M. Masuda T, Kanbayashi N. Fukasawa S, Takeuchi T, Aoyagi T, Umezawa H (1980) Effect of Bestatin on mouse immune system and experimental murine tumors. J Antibiotics 33. 642 9 Leyhausen G. Gramzow M, Steffen R, Zah RZ. Umezawa H, Miiller WEG (1983) Cytochemical indentification of the cell surface bound leucine aminopeptidase, the target enzyme for the immunostimulant Bestatin. J Antibiotics 36, 728 10 Math6 G (1989) Biologic and/or clinical observation on Bestatin therapy and/or prophylaxis and ageing immuno-deficiency and spontaneous tumor appearance. Proc 17th Intern Congress Chemoter, Jerusalem, p 41 II Mathi G, Blazsek I, Canon C. Gil-Delgano M, Misset JL (1986) From experimental to clinical attempts in immunorestoration with Bestatin and zinc. Compl Immurtol
M,
G (1979)
Florentin Restoration
I, Kiger
N.
of impaired
Schulz
J,
immune
functions of aged animals by chronic Bestatin treat-
ment. Imrmrnology 38, 75 Florentin 1, Kiger N, Bruley-Rosset M, Schulz J, Matht G (1978) Effect oi seven immunomodulators on different types of immune responses in mice. In: Human Lymphocyte Differentiation. North Holland, Amsterdam, p 299 Florentin 1. Math6 G, Bruley-Rosset M (1984) Do tuftsin and Bestatin constitute a biopharmacologic immunoregulatory system ? III: Progress in Cancer chemo-immunotherapy (Mathi G, Umezawa H, eds) J Antibiotics Res Ass. Tokyo, p I80
Microbial
Infect
Dis
9, 241
I2 MathC G, Umezawa H, Misset JL, Brienza S. Canon C, Musset M, Reizenstein P (1986) Immunomodulating properties of Bestatin in cancer patients. A phase 11 trial. Biomed & Pharmacoth 40. 379 I3 Ota K. Kurita S, Yamada K. Masaoka T, Uzuka X, Ogawa N (1986) Immunotherapy with Bestatin for acute non lymphocytic leukemia in adults. Cancer Immunol
Bruley-Rosset Math6
wrocka E (1981) Tuftsin and D-arg3-tuftsin possess analgesic action. Experientia 37, 76
Immunoth
23, 5
14 Ota K, Ogawa N (1990) Randomized controlled study of chemoimmunotherapy with Bestatin of acute nonlymphocytic leukemia in adults. Biomed Pharmacother
44, 93
15 Talmadge JE. Black PL, Pelus LM, Abe F (1990) Hematopoietic and hematologic properties of Bestatin in normal and cyclophosphamide myelosuppressed mice. Biomed Pharmacother 44, 85 I6 Schwartz JC, Cros C (1984) In: Bestatin and enkephalins (Math6 G, Umezawal H, eds), p 243 I7 Umezawa H, Aoyagi T, Suda H, Hamada M, Takeuchi T (1976) Bestatin. an inhibitor of aminopeptidase B. produced by actinomycetes. J Antibiotics
29, 97
49
Editorial
estatin, an aminopeptidase with a multi-pharmacological
inhibitor function
G Math6 lnslitur du Cattccr et d’/tttatuttoji~nnPtique 14-16 avenue Paul-Vaillanr-Couturier. 94800 Villejuif,
France
Summary - Bestatin, the dipeptide N-1(2& 3R)-3-amino-2-hydroxy-4-phenyl butanyl) L-leucine is a leucine aminopeptidase and aminopcptidase-B inhibitor. It exerts a direct stimulating effect on lymphocytes (and monocytes) viu its fixation on cell surface leucinc-aminopeptidase. and an indirect effect on monocytes (and lymphocytes) via aminopeptidase B inhibition of tuftsin catabolism. Thus it is an immuno-modifier as shown by the Japanese and also our groups: a) immunomodifier, especially stimulator, in normal young mice; b) immunorestorator and spontaneous tumor preventive agent in aged mice; c) immunorestorator in the elderly and in cancer patients and HIV infected subjects. It stimulates granulocytopoiesis and thrombocytopoicsis irt vim. and can reslore them in viva in myelo-hypoplastic man. The anti-aminopeptidase action of Bestatin also protects cnkephalins againt their catabolism, which encouraged us. with good preliminary results, to study its analgesic action and search for a preventive effect on “desaddicted” toxicomaniac relapses. Bestatin / anti-aminopeptidase
I pharmacological
action
KCsume - La Bestatine, un inhibiteur d’aminopeptidase avec un r6le multi-pharmacologique. Bestutine. lc dipepride N-[(2S, 3R)-3-rttttitlo-2-hydro.vy-4-phcnyl huronoyl) L-leucine est rut inhibireur de la leucine atninopepridase ef de l’antinopepridasc-B. II esis~c 1~1 cjjerl srimulanr direct sur lcs lyttlpltocytes (et les ntonocytes) par i’interttt~diaire de su jkarion sur lu Ieucitte attrittoltc~l)rirktsc de IN surface cellulaire. et utt effer indirect sur lcs ntonocy~cs (er les Iytttphocytes) par I’inrerttrklittire de I’initibiriott du ccmholisnre de la tufrsinc pm I’aminopcp~idase B. La Bestarine esr un ittuttut~o-tnodulant que sitt~uli~retnenr des groupes jopotwis et le tldrrc on1 tttotrtrL’ se cotttplircr enwe aulre cotnme : a) un inununotnodificateur, un srimulateur, cite: dcs souris jeunes et normales: h) un iomntltnorestualtralettr et parall&wrcnt un agent prfventif de la sur\*entte des tutncttrs spotlrutties cite,- lcs souris dgPes: c) rtn inunut~orestaurateur chez des ituntains tigt%, cites des patients rtrk+trs dc cwtcers CI cite: des sujers ittfech par le HIV. II stintrtle la ~mttulocytopoiPse et la rrombocytopoi2se in vitro CI peur IN res~aurer in vivo clre-_ des myE’lo-kypoplnsries imttraines. L’action anti-aminopepridase de la Besturine pro&e ttrtssi lcs ettk~pltalittes comre leurs cataholistncs, ce qtri nous a conduit, avec des rhtl~als pr~linrinaires encotrra,~eatt~s. d itudier /cur urtiott ~muI~~Csiqrte et Li recherciter leurs Cvenruelles capaciks de pre’renir les rechures dc toxicotnanes prhIcrbientcnr soutttis ti uttc cure de dhintoxication. Bestatine / anti-aminopeptidase
inhibiting action gives a only a potential but a very likely chance of curing several, sometimes unexpected and physiologically very different actions. Possessing
pharmaceutic
an
enzyme
/ action pharmacologique
agent,
A peptidase inhibitor
not
Bestatin is a peptidase inhibitor, a small molecular weight dipeptide, N-[(2S, 3R)-3-amino-2-hy-
50
G Mathi:
droxy-4~phenyi butanoyl] L-leucine (fig 1) which was extracted from Sr~q~oq~~s olivoreti [ 171. It was discovered by Umezawa [ 171 who also discovered that it is an inhibitor of leucine aminopeptidase {which is known to bind at the cell surface [9]) and aminopeptidase B (fig 2). The first inhibiting action (or one of the) is the direct immuno-active effect lymphocytes. The second inhibiting action, on aminopeptidase B, works by inhibiting the catabolism of tuftsin [6] (which results from immunoglobulin under the effect of carboxypeptidase [6]), tuftsin which is itself an immuno-(mainly monocytic-macrophage) stimulator.
*--.
STIMULATORY
EFFECT
Fig 1. Chemical s~ruc~urc of Bestatin. (k)-N-[(?S. 3R)-3amino-7-bydrory-4-phenylbutanoyi] t&wine (MW: 308.38).
PEPTWASE tNHlBiTORS BESTATIN AND PEPSTATIN FROM lg
BESTATIN
TUFTSIN H-Thr-Pfe-Arg-Lys-OH
=
INHIBITS TUFI’SIN
\ PEPSlATIN INHWTS \
INHIBITOR Lys-Pro-Arg \
b lo,,g* loopg”
MOUSE INHIBITORY EFFECT r->
-L&INE AMINOPEPTIDASE
Fig 2. Bestatin is: a)aminopcptidase B inhibitor. b) as ppstatin. a leucinc aminopcptidase inhibitor (1). and c) a tuftsin catabolism inhibitor.
An i~~~~o-~od~iato~ statin
and r~sto~ator Be-
it appeared that Bestatin was able to globally stimulate the normal immune functions in young mice (fig 3) [S, 81. More interestingly, we observed that it was able to restore these functions on age ~mmunodepressed mice (fig 3) 141. It not only stimulates in normal mice, T-lymphocyte functions, especially CD4+ interleukin 2
Fig 3. a. Action of Bestatin in the main immunologic functions in young mice. b. Effect of ageing on main immunologic functions. c. Effect of Bestatin according to dose on the immunologic functions in old mice.
Bestatin. an am~no~ptidas~ inhibitor
5
3
i
51
DAY AFTER TREATMENT Fig 4. Effect of oral administrationin mice of Bestatin on 3 conseculivc days on IL-I and IL-2 production by macrophages and spleen cells respectively. 1L-1 and IL-2 pr~uctions, induced irt vitro respectively by LPS and Con A. are amplified 3 and 5 days after a 3-day treatment with Beststin,
1
3
Fig 6. Effects of phospnulipase At (PLAZ) injection on the chemoluminesccnce response of peritoneal cells from immunostimulated mice. Groups of 10 mice were given, on day-a with Bestatin (5 p&mouse ip) and with or without ip injection of 0.7 units PLA?, mouse 30 min before cell harvesl. Each peritoneal cell suspension was tested in~viduatiy. The results are expressed as mV/min f SD.
5
DAY AFTER TR~TMENT Fig 5, Respective effects of Bestatin and amastatin oral administratians in mice for 3 consecutive days on peritonerl macrophage chemolumincsccnce triggered by opsonised zymosan. Fig 7. Pe~toneal cell c~moluminescence after (100 pg) tuftsin and or (25 pg) iv administration.
production (fig 4), but also B cell-antibody production (fig 3a) [5J. It can activate the mono~ytic-ma~rophagic system, increasing interleukin 1 production [fig 4) and peritoneal macrophage chemoluminescence induced zymosan (more intensely than amastatin) (fig 5), and phospholipase A2 (fig 6) or activated by tuftsin (fig 7) 13, 101. In man, it has increased, in cancer patients and NIV infected subjects, the CD&/CDS+ ratio (fig 8) 1121.
Bestatin
More interestingly, Bestatin is able to increase the CD4+ absolute number in the elderly, if the CD8+ cell absolute number is normal (fig 9) [ 101. It can aiso decrease at a certain dose (15 mgld) the number of CD8+CR3~lymph~ytes considered as suppressor cells, while it may increase them at a higher dose (30 mgld) {fig 9) [lo].
G Math6
52
Curative and preventive effects on tumors Bestatin has been shown to exert a regressive effect on murine grafted tumors f I]. It also works in combination with zinc gluconate [ll] as laro semu adjuvant therapy, but only when applied both as stricto-sensu adjuvant and neo-adjuvant treatment to surgery (before and after) (table I) [2]. In addition, when applied between the 16th and the 28th month, it prevents, especially at a high dose, the appearance of spontaneous induced tumors related to ageing, while, as seen above, it restores immunity and delays mortality (table II) [41. In man, clinical controlled trials have been conducted, of which the most interesting has concerned acute myeloid leukemia: the patients being stratified according to age [ 13, 141. There was no increase by Bestatin active immunotherapy of the ABSOLUTE NUMBERS
ABSOLUTE NUMBERS
1800 t
-..._._._._. ,___,_._.__.... ..1.”
(CD4 + , N) (CD8+,
CDB+ NKHi t
NKHt (NKH1 t N)_
Pii: fp. Effect of Bestatin according to dose on immunologic parameters, normality or decrensc.
Table 1. Proportion OF cured animals at day
.._
ml+
CD8t
N)
.
60 after surpy
especially on CD4+lymphocytes according to CDS+ cells
alone or
followed and/or preceded by Bestatin and zinc
iinnlullotherapy, I -0HP cfleinothcrapy or D-Trp-bLH-RH ho~onothe~py. -. Trca~fmW ahe Surgery Zinc gluconate + Bestatin
ffcoadjwmt
Adjutant
Neoadjrrvarrt + Adjtrvattt
0 0
0
6
21
I-OHP(l9)
40
62
66
43
D-Trp-6-LH-RH (20)
10
55
40
50
Bestatin. an aminopeptidase
inhibitor
53
Table II. Prevention by Bestatin of the incidence of spontaneous tumors appearing in FI (C57Bu x BALB/c) micebetween the 16th and 28th month of age. Untreated qed group
Malignanl lymphoma Hepatoma Leukemic lymphoma
Histological type of the tumors No of tumor bearing animals* Total No of animals per group *P
tO0 ~18bestatin-treated aged group
6 1 1
Lymphoma
$=36%
1
+)=5%
= 0.01
.
I
rate which included a rather high proportion of patients younger than 40 (and even 50). The survival rate among controls over the age of 40 is, on the contrary, low, and is elevated by Bestatin at the level of that of patients under 40 years of age [ 13. 141.
Fig 10. Synergy between rhGM-CSF and Bestatin in hemalon cultures.
survival
A pan-hematopoietic storator
stimulator
11
0 6000
and re-
The increase of in viva mice granulocyte number following chemotherapy and radiotherapy has been noted by Talmadge [IS]. We have discovered in vitro, on human CFUs, that Bestatin potentiates the effect of CFS (fig 10) [lo], and we have registered irt vivo in man, this restoration effect on granulocytes and platelets (fig 11) [lo].
150
0
Fig 11. Effects of Bestatin (applied alone to a patienl with subacute marrow aplasia after a long application of cytostatic chemotherapy).
G Math6
54
‘Enkeghalinase’
BESTATIN
lntraneuronal enkephaline
TH~OFW-WJ
>
Aminopeptidases
Release
Fig 12. Bestarin inhibits endogenous enkephaline catabolism via aminopeptidase inhibition.
Other effects, especially psychic
7 Herman ZS, Stachura Z, Opiela L. Ziemion IZ, Na-
that enkephalins are pentapeptides of the general formula tyrosine-glycine-glycinephenylalanine, the last amino acid being either methionine or leucine. Bestatin inhibits at least three aminopeptidases in the brain (fig 12) 116) and tuftsin is known to exert an analgesic effect [7]; we have searched for: a) an analgesic effect: b) a relapse inhibitory effect in toxicomaniacs after desaddiction. Our studies, which are in phase II progress, are very encouraging. It is known
References Abe F. Shibuya K, Uchida M, Takahashi K, Horinishi H, Matsuda A, lshizuka M, Takeuchi T, Umezawa H (1984) Effect of Bestatin on syngeneic tumors in mice. Gunn 75, 89 Bourut C, Chenu E, Math6 G (1988) Management of Minimum
Residlral
Malignancy
in Man.
Oxford, p 59 Bravo-Cuellar A. Homo-Delarche
Perga-
mon,
bouys S (1990)
Phospholipase
AZ,
F, Orbach-Aran in vivo im-
munomodulator. Prost Leltko Essent Fatty Acids 40, 31
8 Ishizuka M. Masuda T, Kanbayashi N. Fukasawa S, Takeuchi T, Aoyagi T, Umezawa H (1980) Effect of Bestatin on mouse immune system and experimental murine tumors. J Antibiotics 33. 642 9 Leyhausen G. Gramzow M, Steffen R, Zah RZ. Umezawa H, Miiller WEG (1983) Cytochemical indentification of the cell surface bound leucine aminopeptidase, the target enzyme for the immunostimulant Bestatin. J Antibiotics 36, 728 10 Math6 G (1989) Biologic and/or clinical observation on Bestatin therapy and/or prophylaxis and ageing immuno-deficiency and spontaneous tumor appearance. Proc 17th Intern Congress Chemoter, Jerusalem, p 41 II Mathi G, Blazsek I, Canon C. Gil-Delgano M, Misset JL (1986) From experimental to clinical attempts in immunorestoration with Bestatin and zinc. Compl Immurtol
M,
G (1979)
Florentin Restoration
I, Kiger
N.
of impaired
Schulz
J,
immune
functions of aged animals by chronic Bestatin treat-
ment. Imrmrnology 38, 75 Florentin 1, Kiger N, Bruley-Rosset M, Schulz J, Matht G (1978) Effect oi seven immunomodulators on different types of immune responses in mice. In: Human Lymphocyte Differentiation. North Holland, Amsterdam, p 299 Florentin 1. Math6 G, Bruley-Rosset M (1984) Do tuftsin and Bestatin constitute a biopharmacologic immunoregulatory system ? III: Progress in Cancer chemo-immunotherapy (Mathi G, Umezawa H, eds) J Antibiotics Res Ass. Tokyo, p I80
Microbial
Infect
Dis
9, 241
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