JPS6183185A - Metal phephorbide derivative and metal porphyrin derivative - Google Patents

Abstract

NEW MATERIAL:A compound of the formula [R1 is CHCH2, CH(OH)CH3, CH (OCH2CH2OH)CH3, R2 is CH3, CHO, CH2OH; R3 is CO, OH, OCOCH3; R4 is H, CO2CH3, CO2CH2CH2OH; R5 is CO2H, CO2Na, CO2CH3; M is Si, Ga, In, Pt; X is halogen, hydroxyl, pyridien]. EXAMPLE:Si-9-Desoxo-9-hydroxoypheophobide. USE:Diagnostic or remedy for cancer; it has high affinity to cancer and properties of destructing cancer cells with electromagnetic waves without phototoxicity. PREPARATION:For example, a metal complex, e.g., Si Ga or Ge is obtained by dissolving a phophorbide derivative such as 9-desoxo-9-hydroxy-pheophorbide in pyridine and adding a metal salt such as SiCl4 to the solution with heat under stirring, while In or Pt complex is formed using acetic acid as a solvent.

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Industrial Application Field The present invention provides a novel cancer treatment system that uses a metal complex of pheophobite and porphyrin derivatives as an active ingredient for diagnosis, marker, and missile therapy. Regarding formulations.

(b) Conventional technology The present inventors have been involved in research on chlorophyll-related compounds for a long time, and the first method to obtain pheophornoids is to use photosynthetic organisms that have a cheap and stable source of raw materials, which is economically efficient. Established a method for separating pheophobite (Takumi Kokai Sho 58-69884).

Patent Application No. 58-198934, in which the present inventors discovered for the first time that plant-derived heptagonal ophobide derivatives have the property of selectively binding to cancer tissues and reacting when exposed to the source.
(filed on June 22, 1982), -! Blood-derived porphyrin derivatives have been known to have these properties. However, it was not thought that gold-spear-coordinated pheophobite and porphyrin derivatives had a concentration against cancer and no response to light.

The pheophorbite compound, which was first applied by the present inventor, is a phorbin derivative derived from plant chlorophyll (Japanese Patent Application No. 58-198934, 1989-6-22, previously applied substance), and the porphyrin compound, which is animal hemoglobin. It is known that porphine derivatives derived from porphine have binding to cancer tissues and have optical chirality.

Practical application for cancer treatment is underway.

Related compounds of gold-h pheophobite derivatives and metal porphyrin derivatives (hereinafter referred to as the present substance) include animal-derived hemin (FeR form of porphine), plant-derived resin oophyll (Mg complex of forpine), Cu and F.
e Chlorophylrin (Cu and Fe complex of 7 orbin), CO-protoporphyrin (C of porphine).

Na body), Hg-porphyrin (Hg complex of porphine) [Tasaki et al., Cancer Chemoth
erapyReports, 13.41 (1961
)] etc. In addition, in recent years, Futoshi et al. has reported a method for producing Ge-protoporphyrin (Japanese Patent Publication No. 57-31688), and Unno et al. has reported a method for producing a hematoporphyrin Si derivative (Japanese Patent Publication No. 58-201793).

However, hemin is a blood component, and chlorophyll is a plant component C.
u and Fe Chlorophyllin is a food additive coloring agent and deodorizing agent, and these drugs have no cancer affinity. CO-protoporphyrin had been approved as an anticancer drug until recently, but it was rejected by the Pharmaceutical Affairs Council (August 10, 1981). Although Hg-porphyrin derivatives are also recognized to have anticancer effects, Hg
g is highly toxic and is unsuitable as a medicine. Regarding Ge-protoporphyrin, it is stated that it has an effect of improving hepatic dysfunction as well as a side thread effect, but the data has not been published, and there is no mention of its selective accumulation in cancer cells. Not yet. Hematoporphyrin Si
The derivative is a metal compound in which the St metal is not coordinated to the nitrogen atom of the porphyrin skeleton that we are aiming for, but the St metal is directly bonded to the side chain of the skeleton, and therefore has phototoxicity.

(c) Problems to be solved by the invention Pheophorite and porphyrin compounds are useful substances that have selective accumulation in cancer cells, reactivity to light, and cancer destruction. On the other hand, photodynamic effects act as phototoxicity, which increases the difficulty in actual treatment. That is, cancer patients have to live in the dark for 48 to 72 hours after administering these drugs, which is extremely painful. In addition, some patients may develop phototoxicity even 1 to 2 weeks after administration due to the high cancer accumulation properties of these compounds, which is why these compounds cannot be used to facilitate diagnosis. Ta.

The present invention aims to eliminate the phototoxicity of these compounds and obtain a new cancer therapeutic drug 1 diagnostic agent.

Therefore, the present inventors discovered that these pheophorbites and porphyrins! Let the four bodies be metal complexes.

We believe that eliminating phototoxicity may open up the possibility of application as a new cancer treatment drug and diagnostic agent.

As a result of extensive research, we have found that precipitate substances and porphyrin derivatives such as Mg, St, Ni, Zn+ Ga, Ge
, Innpt and other metal complexes have a remarkable tendency to accumulate in cancer cells.

It has been found that this metal complex compound has pharmacological effects such as strong reactivity to external energy and destructive action, and on the other hand, this metal complex compound is extremely superior because it does not have the photothermogenic properties of pheophobite and porphyrin derivatives.

Next, a method for preparing the compound of the present invention will be explained.

General formula (]), (21) The method for producing the substance of the present application is to first adjust the side chain of the phorbin skeleton according to the method for producing the preformed substance, and then to prepare the preformed material k S I C141G a C
1s +GeCl4. InC15 Kamata n K2P t
5i-9-desoxo-9-)・
Hydroxypheophobide (hereinafter referred to as I) +
Ga-2-desethynyl-2-(1-acetyloxyethyl)-pyropheophobite (hereinafter referred to as ■),
Ga-ethylene glycol sea 10 b-7 eophopate (hereinafter referred to as So) + Ge -2-desethynyl-2-(1-hydroxyethyl)-9-nasoxo-9-hydroxypheophopide (hereinafter referred to as I)
V), In-monoacetylenalene glycol, mono-10b-pheophorbate (hereinafter referred to as ■),
pt-7 eophopide (hereinafter referred to as v1) respectively.

Implementation of the present invention + 811 (J a + C
For metal complexes such as r e , the precipitate was dissolved in pyridine, and then each gold 14 salt was added and reacted with heating and stirring. Obtain each IV. Regarding the metal complexes such as In and Pt that I received, I bath-dissolved them in nF acid according to the previous application, and then heated them in step 4. ) Add each metal salt to react while stirring V,
These reaction endpoints can be easily identified using thin-layer chromatography or an ultraviolet handscope.

In addition, the present substance of general formula (3) can be obtained by first treating the 11111 button of the porphyrin skeleton with hydrobromic acid or acetylating the hydroxide obtained after the treatment in the same manner as obtaining the front axis substance. Prepare each oxide or acetylated product.

Next, these obtained compounds were mixed with Mg(C10J2+
GaC15+ GeCl4. Ni(OAc)t
+ Zn(OAc)z.

Treated with I nC13 or Eu(AA)s, M
g-di2.4-(1-ethanealcoholoxyethyl)-deteroporphyry/(hereinafter referred to as V1).

Ga-di2,4-(1-ethanealcoholoxyethyl)-deuteroporphyrin (hereinafter referred to as vllll)
, Ge-di2.4-(s-ethaneacetateoxyethyl)-deuteroporphyrin (hereinafter referred to as ■)
, Ni-hemadoporphyrin (referred to as 9-lower X), Z
n-di2.4-(1-ethanealcoholoxyethyl)-dez-telopornyrin (hereinafter referred to as
-Hemadoporphyrin (hereinafter referred to as "S") and Eu-G2,4-(+-ethanealcoholoxyethyl)-deuteroporphyrin (hereinafter referred to as "■") are produced.

In carrying out the present invention, + for metal complexes such as Mg, Ga, Ge, etc., each porphyrin derivative is dissolved in pyridine, and each metal salt is then added and reacted with stirring under heating to obtain compounds (1) to (2), respectively. Ni.

For metal complexes such as Zn and In, each porphyrin conductor is dissolved in acetic acid, and each metal salt is added while heating or pressing to react to obtain X-Xll. For metal complexes such as +Eu, each porphyrin derivative is dissolved in trichlorobenzene, and then Eu(AA)3 is added and reacted with heating and stirring to obtain (2).

Another method for producing the substance of the present application is to form a metal complex at the stage of starting from the raw material methylpheophobite or dimethylprotoporphyrin. It is also possible to obtain favorable results. The combination is not limited to this, as long as it is a hydroxylation reaction, hydrolysis reaction, esterification reaction, or metal complexation reaction.

It is also preferable to add an antioxidant, a solvent, etc. and heat, but this is not an essential condition.

As for starting materials, pheophorbate is a typical plant-derived material, and protoporphyryf is a typical animal-derived material, but the starting materials are limited to these if they are obtained from photosynthetic organisms or blood components. isn't it.

The ultraviolet absorption spectrum data of the substance of the present application are shown in Table 1, No. 1.

Table 1 of Table 1 shows the nm of maximum absorption (λmax) of each substance.

Porbin skeleton Porphine skeleton (e) Actions Next, we will explain the pharmacological action of the claimed substance.7 The claimed substance is selectively concentrated in cancer tissues, is slowly excreted from cancer cells, and does not react even when exposed to light.

External energy other than light, ie microwaves.

When irradiated with electromagnetic waves or the like, it reacts and is excited to generate singlet oxygen, which has a strong oxidizing effect, and the action of this -quaternary oxygen destroys cancer cells. Furthermore, it should be promptly excreted from normal organs and cells, so it will not harm normal cells.

7Almost most of the eophapeids and porphyrin derivatives have strong effects on light.

By forming both of these derivatives into metal complexes, the advantageous property of selective accumulation in cancer cells can be maintained, while the disadvantageous strong effect on light can be eliminated. Therefore,
Metal complex derivatives are used as diagnostic agents because they do not exhibit phototoxicity.
1141T tumor marker, effective for missile therapy.

This substance is a useful substance that possesses all of these properties (cancer affinity, non-phototoxicity, and cancer-destructive property against external energy).

(f) Examples The pharmacological effects of the substance of the present application and back-door methods will be explained below using examples.

Example 1 Laser irradiation in excised organ (excitation fluorescence spectrum) 14-2 transplanted with nitrosamine carcinogenic rW5 cancer cells
The test drug Ga-monoacetyl ethylene glycol mono-10b-pheophorate sodium salt, Mg-G2 diluted with physiological saline (1 m/g) was given to Golden hamsters (group of 5 animals) on the first day. .4- (1
-ethanealcoholoxyethyl) -disodium of deuteroporphyrin IIvlD"a,Ge -G2
, 4-(1-ethaneacetate ox7ethyl)-deuteroporphyrin (2) disodium salt + In
- After each of the disodium salts of hemadoporphyrin (2) was injected intravenously (l), cancer cells and other organs were removed, and Nt
-pulsedlaser(Nt+ 337 nm
, 2ns 400-11000n),
The excitation fluorescence spectrum was measured and 470nrfl NAD
Wavelengths of 600 to 900 nm were investigated using the peak wavelength of H as a reference.

Figure 1 shows that each drug significantly accumulated in cancer cells after 24 hours, but did not remain in other cells.

In addition, when similar tests were conducted on other drugs, Table 1
The results shown in Table 2 were obtained. Table 2 of Table 1 shows the excitation fluorescence spectra of each organ extracted 24 hours later.
600 using the NADH peak wavelength of 70 nm as reference 1.
The peak wavelength tX at ~9001 m was obtained (If O) As is clear from the above results, these Kanasu porphyrin-related compounds have a remarkable selective affinity for cancer cells. 2 Dissolve 9-desoxo-9-hydroxypheophobide 12 in 90jfj of pyridine, then add 5ict
, o, sp (dissolved in pyridine 101R1), heated to 165° C. in a sealed tube, and reacted for 2 hours. The end point of the reaction was determined using thin layer chromatography and an ultraviolet ray scanner. After the reaction, the reaction solution was added with IQQdl of water and extracted with CHCl s. CHCj s layer gold wash,
After drying, the mixture is concentrated to obtain red St-9-desoxo-9-hydroxypheophonoid (1) (yield: 0.7 y). This yield is 60%.

The obtained I was dissolved in methanol and neutralized by adding a 5% NagCOs aqueous solution to obtain the sodium salt of I. This yield is 9
It is 8chi.

Example 3 1 g of 2-desethynyl-2-(1-acetyloxyethyl)-pyropheophobite was dissolved in 90% of pyridine, and then GaC1,o, s2 (pyridine 10j17
Add solution #) to the solution, heat to 160°C in a sealed tube, and react for 1 hour. Thereafter, the same operation as in Example 2 was carried out to obtain blue-green Ga.
-2-desethynyl-2-(1-acetyloquinethyl)-bilofehopide (n) 'e obtained (yield 11
). This yield is 82%.

Example 4 Ethylene glycol di-10b-7 eophobe) IP
'iDissolved in 90% pyridine and then 30% GaCA'.
Add 59 (dissolved in pyridine tomg) to 160 in a sealed tube.
Heat to ℃ and allow to react for 1 hour. Hereinafter, in the same manner as in Example 3, blue-green Ga-ethylene glycol-di-1 was obtained.
Obtain 0b-pheophopate 01) (yield 0.99)
This yield is 73%.

Example 5 2-desethynyl-2-(1-)-idroxyethyl)-9-desoxo-9-idroxypheophoba rdo1y was dissolved in pyridine 9Qmz and then G
eCt, 0.59 (P9F19) and reacted for 2 hours. The same procedure as in Example 3 was carried out to convert red Ge -2-desf= to -2- (
1-hydromioxyethyl)-9-desoxy-9-
Hydroxypheophobide GV) is obtained (yield 0.BS+), which yield is 66%.

Example 6 Monoacetyl ethylene glycol mono-10b-7 eophopate 1y was dissolved in acetic acid 90% and then In
C/3 0. sy (MV dissolved in 10 ml of acetic acid)
, NaCJ O. 5 yf addition, t addition flPr return (At. L.

Allow to react for 1 hour. The end point of the reaction was determined by thin layer atomography and ultraviolet light. After the reaction, 100 ml of water was added to the reaction solution and extracted with CH2C/.

The CH,Clt layer is washed with water, dried and concentrated to obtain green In-monoacetylethylene glycol mono-10b-7 eophopate (V) (yield xp).

This yield Vi is 80%.

Example 7 Pheophorite 11 was dissolved in 9sdKyB acetate, then KsPtClp, 0. ．． 59 (Dissolved in 5 d of water) Add acetic acid powder 19, heat to reflux, and react for 2 hours. Thereafter, the same procedure as in Example 6 was carried out to obtain pt-7 eophobide (yield: 1.39). This yield is 85 cm.

Example 8 GZ, 4-(x-ethanealcoholoxyethyl-deuteroporphyrin IPi pyridi 7100*t) was dissolved, then ivIg((jo4)t059 was added, heated to reflux, and reacted for 1 hour. Example 3 below. In the same manner as above, a pale pink -di-24-(1-ethanealcoholoxyethyl)-deuteroporphyrin (VID) was obtained (yield @19).

This yield Vi is 92%.

Example 9 Di-14-(1-ethanealcoholoxyethyl)-deuteroporphyrin 1y was dissolved in 90ml of pyridine, and then GaC/0.5 F (dissolved in pyridine IOd) was added and heated at 160°C in a sealed tube. and react for 1 hour. Thereafter, the procedure was repeated in the same manner as in Example 3 to obtain crimson Ga.
-Z4-di(1-ethanealcoholoxyethyl)-
Deuteroporphyrin ■ is obtained (yield: 1.19).

This yield is 90%.

Example 10゜G24-(1-ethaneacetateoxyethyl)-deuteroporphyrin 19 was dissolved in 90jL of pyridine, 9 then GeC1,0,5y (dissolved in 1Qal of pyridine) was added, and the mixture was heated to 180°C in a sealed tube. Heat and react for 2 hours. Hereinafter, in the same manner as in Example 3, pale pink Ge −
Di2.4-(1-ethaneacetateoxyethyl)-
obtain dataroborfiri/(2) (yield o, s
y). This yield is 67%.

Example 11 Hematoporphyrin 15I was dissolved in 90ml of acetic acid, and then N1(OAc)tO,5F (dissolved in IQad acetate) and 0.52ml of sodium acetate were added, heated to reflux, and reacted for 1 hour. After that, perform the same operation as in Example 3 to obtain red N.
i-hemadoporphyrin (1) is obtained (yield t l
y). This yield is 85%.

Example 12゜G2.4-(1-ethanealcoholoxyethyl)-
Deuteroporphyrin dimethyl ester 19 (:)
i(j39 dissolved in OSIP and then Zn(OAc)
To the mixture were added t o and S P (dissolved in 10 ml of methanol), heated to reflux, and allowed to react for 1 hour.

Thereafter, the same procedure as in Example 3 is carried out to obtain a Zn@ body. The obtained complex was hydrolyzed with an alkali using a conventional method to obtain Zn-di-24-(1-ethanealcoholoxyethyl)-defoporphyrin (yield: 0.9 p), with a yield of 73%. be.

Example 13 Hematoporphyrin 12 was dissolved in 90d of acetic acid and then InCj30. s y (10 MIVC bath solution in vinegar)
Then, acetic acid powder was added thereto, heated under reflux, and reacted for 1 hour.The following procedure was repeated as in Examples 16 and 1[1j] to obtain In-hemadoporphyrin~Df (Yield IO, 7p). This yield was 51%.

Example 14 λ4-di(neethane alcoholoxyethyl)-tuteloporphyrin dimethyl ester 19i Trichlorobenzene 10 (1/9) dissolved in 9 then En(AA),
Add o, sy and heat to reflux, and leave to cool for 2 hours. The obtained reaction solution was mixed with Kei! The mixture was subjected to 4:'2 column chromatography, firstly trichlorobenzene was removed with n-hexane, then the target product was eluted with ethyl acetate, and then concentrated. The obtained Eu complex is hydrolyzed with an alkali in a conventional manner to obtain red Eu-2,4-di(1-ethanealcoholoxyethyl)-deuteroporphyrin (■) (yield: 1). This yield is 65 cm.

(G) Effects of the Invention The compounds of the present invention have pharmacological effects such as accumulation in cancer cells, reactivity to external energy, and destruction of cancer cells, and have no reactivity to light. This compound is different from conventional pheophobite and porphyrin derivatives, and is extremely useful as a cancer diagnostic FIF+ drug.

The present invention is a metal complex of a porphyrin derivative in a broad sense,
Both porphyrins (in a broad sense), such as plant-derived chlorophyll (phorbin derivatives) and animal-derived hemoglobin (porphine derivatives), which are inexpensive and stable sources, can be used as raw materials.

The first substance in this exhibition is a metal volphyri/(forbin and porphine) related compound that has characteristic physiological activities such as cancer affinity, non-phototoxicity, and action against external energy V, so it can be used as a cancer diagnostic agent, an AJ remarker, and other substances. It can be used directly as a medicine for missile therapy as a carrier for anticancer drugs, or has almost unlimited uses as a raw material for their production, and can be produced easily and in a short time.
It is a highly economical and extremely useful substance.

[Brief explanation of drawings]

Figure IIfi shows the excitation fluorescence spectra of each organ 24 hours after intravenous injection of each drug. Z4- (
Figure 3 shows the disodium salt of Ge-G2.
Disodium salt of 4-(1-ethaneacetateoxyether)-deuteroporphyrin (Treasury), Figure 4 FiI
FIG. 2 is a chart regarding the affinity for cancer cells when using each drug of the disodium salt of n-hemadoporphyrin (6). FIG. l Physiological saline (control) 2, tumor 3, liver 4 kidney 5 serum

Claims (3)

[Claims] (1) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (1) In the formula, R_1 is CH=CH_2, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ or ▲
There are mathematical formulas, chemical formulas, tables, etc. ▼ R_2 is CH_3, CHO or CH_2OHR_3 is C
=O, OH or OCOCH_3 R_4 is H, COOCH_3, COOCH_2CH_2
OH or COOCH_2CH_2OCOCH_3R_5
is COOH, COONa or COOCH_3M is Si,
Ga, Ge, In or Pt X represents a halogen atom, OH, pyridine, OCOCH_3 or the like. A metal pheophobide derivative represented by (2) General formula ▲ There are numerical formulas, chemical formulas, tables, etc. ▼ (2) In the formula, R_1 to R_5, M, and X represent the same residues as in the general formula (1). A dimeric metal pheophobide derivative represented by . (3) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (3) In the formula, R_6 and R_7 are CH=CH_2, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ R_8 is COOH, COONa or COOCH_3M is Mg, Si, Ni, Zn, Ga, Ge, In, Eu or Pt X represents a halogen atom, OH, pyridine, OCOCH_3, etc. A metalloporphyrin derivative represented by However, metal complexes of protoporphyrin and hematoporphyrin are excluded.