JPH05302919A - Assay medicine for antitumor effect of host intervening anticancer agent - Google Patents

Abstract

PURPOSE:To make it possible to monitor whether antitumor effect can be expected or not in a clinical way when host intervening anticancer agent is used for a patient by containing lentinan or Sonifiran. CONSTITUTION:This assay medicine contains lentinan or Sonifiran. When the lentinan as the amount of anticancer agent is administered, blood-vessel permeability rises up. Then, the macrophage in the organism is activated by the administering of the lentinan, and the second path of the complement is also activated. The complicated biological reaction caused by various freed mediators occurs. As a result, the blood-vessel peremeability is changed. It is estimated that the occurrence of the reaction such as this at the local tumor position is related to the apperance of the antitumor effect. Therefore, the antitumor effect can be judged by inspecting the change in reaction in tumor sample, which is obtained by surgically extracting the same reaction and the change in intracutaneous reaction of patient.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a drug for determining the antitumor effect of an antitumor agent, and more specifically, it provides a test agent for clinically determining the expression of the antitumor effect of a host-mediated antitumor agent. ..

[0002]

2. Description of the Related Art Various anticancer agents used for the treatment of cancer patients are known which have different chemical structures, action mechanisms and the like. Among these drugs, the so-called host-mediated anticancer drug that exerts an antitumor effect through activation of immune function, which is a defense mechanism of the body, is superior to chemotherapeutic drugs based on selective toxicity because it has no side effects. There is.

As such an immunotherapeutic agent, many ones such as lentinan, sonifilan, picibanil, krestin, bestatin and the like are known and used as therapeutic agents for cancer patients. Since these are drugs that exert an antitumor effect through activation of the patient's immune system and biological defense system, they are affected by background factors of the patient, and there are individual differences in the expression of the antitumor effect. , The difference in the intensity of action tends to be recognized depending on the clinical condition of the patient.

Therefore, it is difficult to expect an effective treatment using these host-mediated anticancer agents only by judging the effectiveness of these agents on the basis of general clinical symptoms. In order to effectively treat with these drugs, it is desirable to be able to easily evaluate in the medical field whether the patient responds to these drugs at the early stage or before the treatment.

[0005] Further, it is considered that the optimal dose, the number of administrations, the administration time, and the application method of the chemotherapeutic agent to be used in combination with the host-mediated anticancer drug may differ depending on the individual patient. It is desired to develop a method for bedside monitoring of the optimal administration method for each animal.

In order to exert an antitumor effect by a host-mediated anticancer drug, antitumor effector cells such as T cells, macrophages and neutrophils and inflammatory cells are activated, and further these cells infiltrate the tumor local area. It is considered essential to exert a synergistic effect that promotes vascular permeability as possible. However, no attempt has been made to grasp the degree of expression of the antitumor effect from the change in responsiveness of the vascular system at the tumor site.

On the other hand, when lentinan is administered to mice,
Vascular permeability of the auricle is increased and the auricle becomes red (VD
H reaction) has been reported (Maeda et al . Int . J. Imm
unopharmac .6 , 493 (1984)), but this is a phenomenon that can be observed only in white experimental animals and cannot be used as a clinical test method for humans that meets the above purpose without quantification. ..

[0008]

DISCLOSURE OF THE INVENTION The present invention aims to determine whether an antitumor effect can be expected when a host-mediated carcinostatic agent is used for a patient in order to effectively treat the cancer patient with the host-mediated carcinostatic agent. To monitor clinically,
Another object of the present invention is to provide a test agent that enables the selection of a treatment method for optimizing the treatment effect in the medical field.

[0009]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventor has observed that the antitumor effect of a host-mediated antitumor agent monitors changes in tumor locality and vascular responsiveness of skin. The present invention was completed by finding that the evaluation can be performed by

That is, the present invention is a test drug for determining the therapeutic effect of a cancer drug administered to a host-mediated anticancer drug, which is characterized by containing lentinan or sonifilan, and A test drug for determining the therapeutic effect of these carcinostatic drugs on a cancer patient administered with a host-mediated carcinostatic drug, which is an antitumor effect test drug characterized by containing a vascular permeability-enhancing substance.
It should be noted that any substance other than lentinan and sonifiran may be used as the test agent of the present invention as long as it has a polysaccharide having a β-1,3 glucan structure as a main component.

In the present invention, the host-mediated antitumor agent refers to an antitumor agent that exerts an antitumor effect through activation of immune function, and is clinically applied such as lentinan, sonifilan, picibanil, krestin, bestatin, and the like. There are many under development known to have the action of

The present invention will be described in detail below. <1> Antitumor Effect Test Agent Containing Lentinan The antitumor effect test agent containing lentinan of the present invention only needs to contain lentinan, and other components may be used. However, it is desirable that it does not contain components that cause redness on the skin. The dosage form may be liquid or freeze-dried powder. In addition, mannitol, lactose, cyclodextrin and the like can be added as excipients to the extent that they do not irritate the skin.

The lentinan may be used as it is or may be a low molecular weight one having a molecular weight of 1,000,000 to 10,000. Next, a method of using the above-mentioned test agent will be described.

The patient to be administered the host-mediated anticancer drug,
The test drug is administered by intradermal injection 3 to 21 days after the start of the administration. It is desirable to administer several times by changing the administration site at regular intervals.

The site of administration is preferably intradermal or subcutaneous injection on the back or upper arm, but other sites can be selected depending on the patient's condition. When lentinan is used as a test agent, the dose is preferably in the range of 0.05 to 5000 μg,
The dose may be changed and administered to several sites.

The test drug is administered, and changes in vascular responsiveness are monitored after a certain period of time, preferably 20 minutes to 1 day. This monitoring can be performed, for example, by measuring the degree of redness occurring at the test drug administration site or by measuring the skin surface temperature around the administration site by thermography.

When a skin reaction is difficult to be observed, clinically, a dye such as phenolsulfonphthalein injection used for examination of liver function and renal insufficiency, fluorescein sodium used as a diagnostic agent for fluorescence imaging, and the number of lentinan doses are administered. If intravenously injected after a minute, the reaction of the living body is amplified and it is easy to measure.

In addition, for a patient who is scheduled to administer a host-mediated anticancer drug, only the test drug is administered before administration, and the intradermal reaction is observed in the same manner as above, and used as a control index value for the patient, and the administration against this control value is performed. It is desirable to track subsequent changes in values. An antitumor effect can be expected when redness and pigment reaction are remarkably observed, or when the skin surface temperature is high, or when the values of these reactions are higher than the control index value of the patient.

Also, the same effect can be obtained by using sonyfiran instead of lentinan. The dosage form, additives, method of use, and dose may be similar to lentinan. Further, as long as it has a polysaccharide having a β-1,3 glucan structure as a main component as described above, compounds other than lentinan and sonifiran can be used as the test agent of the present invention.

Further, a test kit comprising an antitumor effect test drug containing lentinan or soniphyran and a medical dye phenolsulfonphthalein or sodium fluorescein is also one of the test drugs included in the scope of the present invention.

<2> Antitumor Effect Detecting Agent Containing Vascular Permeability Enhancer In place of the polysaccharide-related substances such as lentinan and sonifiran, a vascular permeability enhancer can be used as the test agent of the present invention. Examples of such substances include bradykinin, kallidin, histamine and the like.

The dosage form, additives, and method of use are the same as those of the lentinan-containing test agent except the amount used. The dose is usually 0.01 for histamine.
ng to 1 ng, 10 in the case of kallidin and bradykinin
^-12 M to 10 ^-6 M is preferable. Of course, the above value is a general index, and may be increased or decreased depending on the patient.

[0023]

[Action]

(1) Action of lentinan Lentinan is a high molecular neutral polysaccharide extracted from shiitake mushroom and has glucose as a constituent component. Although it is used as a carcinostatic agent because it has an immunostimulatory action, in the present invention, lentinan is used as a test agent for determining the therapeutic effect of a host-mediated carcinostatic agent such as lentinan, sonifilan, picibanil, and krestin.

Administration of lentinan as an anti-cancer agent increases vascular permeability. This is because the administration of lentinan activates macrophages in the body and also activates the alternative complement pathway, which causes a complex biological reaction by various medulators released here, resulting in a change in vascular permeability. It is presumed that such a reaction also occurs locally in the tumor, which is involved in the expression of the antitumor effect. Therefore, it is expected that the antitumor effect can be determined by investigating changes in the intradermal reaction of a tumor specimen or a patient surgically removed from the same reaction.

On the other hand, when lentinan is intravenously administered to mice, a VDH reaction (vasodilation and bleeding) is observed in the auricle. This is because administration of lentinan increases vascular permeability and red blood cells leak from blood vessels. This VDH
The reaction is correlated with the increase in the intradermal reaction, and if the change in the intradermal reaction can be monitored, it will provide a method of knowing the degree of antitumor effect expression not only in the white mouse but also in the human medical field. be able to. This correlation is proved in Example 1.

In humans, VDH reaction cannot be observed only by administration of lentinan as an anticancer agent, but when administered as an anticancer agent and intradermally administered with lentinan in a state where vascular permeability is increased, a reaction occurs in a normal state. Even at such a concentration that does not occur, an intradermal reaction such as redness is observed.

(2) Action of vascular permeability-enhancing substance When the vascular responsiveness is changed by the administration of a host-mediated anticancer drug such as lentinan, sonifilan, picibanil, krestin, etc., the sensitivity to the vascular permeability-increasing substance is increased. As a result, when a substance such as histamine or bradykinin is intradermally administered, redness or the like occurs even at a concentration at which no change occurs by itself. Therefore, like the lentinan, a vascular permeability-enhancing substance can be used as a test agent.

The correlation between the increased susceptibility to the vascular permeability-enhancing substance and the antitumor effect in the mice administered with the host-mediated carcinostatic agent is proved in Example 2 and the following. Sonifiran also has the same action as lentinan.

[0029]

EXAMPLES Examples of the present invention will be described.

[0030]

Example 1 An example of a test drug containing lentinan will be described, and the correlation between the VDH reaction and the increase in the intradermal reaction due to the administration of a host-mediated anticancer agent will be shown by the experimental results.

(Method) Lentinan 5 mg / kg as a carcinostatic agent or physiological saline was administered to A / J mice by intravenous injection three times every other day, and 6 times from the first administration.
After a day, the degree of increase in intradermal reaction was observed.

The intradermal reaction was examined by the resulting increase in vascular permeability. For vascular permeability, 300 μl of 1% Evans blue solution was intravenously injected into a mouse, and immediately thereafter 25 μl of lentinan or physiological saline as a test drug of various concentrations was intradermally administered to the lower abdomen, and 20 minutes after the end of Evans blue administration. After that, the mouse was sacrificed, and the area of the pigment spot due to Evans blue formed in the skin was evaluated as an index. The area was measured with a Lussex III image analyzer (manufactured by Nireco).

Hereinafter, lentinan as a carcinostatic agent is referred to as "anticancer drug", and lentinan as a test drug is referred to as "test drug".
On the other hand, the VDH response of the pinna was evaluated by visual observation.

(Results) As shown in FIG. 1, the accumulated amount of Evans blue increased according to the amount of the test agent intradermally administered to the lower abdomen of the mouse. This is because the intravenously administered Evans blue leaked out of the blood vessel by the intradermal administration of the test drug (Fig. 1 ●).

On the other hand, in mice to which physiological saline was pre-administered instead of the anti-cancer drug, no significant leakage of Evans blue was observed even when the test drug was administered intradermally (Fig. 1).
○).

On the other hand, the VDH reaction in the auricle was observed only in the mouse to which the anticancer drug was intravenously administered in advance, and was not observed in the mouse to which the physiological saline was administered.

From these results, VD by administration of anticancer drug
It was found that there is a correlation between the H reaction and the increase in the intradermal reaction, and that the increase in the intradermal reaction can be known by subcutaneously administering the test drug after the administration of the anticancer drug.

From the above, it is understood that the degree of expression of the antitumor effect of the host-mediated antitumor agent can be determined using lentinan.

[0039]

[Example 2] Next, an example of a test drug containing a vascular permeability-enhancing substance will be described, and the experimental results of examining the change in sensitivity to the vascular permeability-enhancing substance in mice pre-administered with a carcinostatic agent will be shown. Bradykinin was used as the vascular permeability-enhancing substance.

(Method) In the same manner as in Example 1, lentinan 5 mg / kg or physiological saline was administered to A / J mice by intravenous injection three times every other day, and 5 times from the first administration.
After a day, the degree of vascular permeability enhancement was observed.

Regarding the degree of vascular permeability, Evans blue was administered by intravenous injection, and immediately thereafter, 25 μl of bradykinin or physiological saline was intradermally administered to the lower abdomen of the mouse.
After 0 minutes, the area of pigment spot was evaluated as an index.

(Results) As is clear from FIG. 2, leakage of Evans blue was observed in mice pre-administered with an anticancer agent when bradykinin was administered after intravenous injection of Evans blue (FIG. 2). On the other hand, the amount of Evans blue leaked out was small even when bradykinin was administered to mice to which physiological saline was administered instead of the anticancer drug (FIG. 2).
○).

From these results, it was found that the administration of anticancer drug promotes the effect of bradykinin on the enhancement of vascular permeability. Further, a VDH reaction was observed in the mice administered with the anticancer drug, but not in the mice administered with physiological saline.

From the above, the antitumor effect of the host-mediated anticancer drug can be determined by observing changes in vascular permeability using a vascular permeability-enhancing substance.

[0045]

Example 3 In order to examine the correlation between the change in vascular permeability and the intradermal reaction as described above, lentinan was administered as a carcinostatic agent to T cell deficient mice, and the sensitivity to bradykinin was examined.

(Method) A normal mouse (ICR mouse),
Using T cell deficient mice (ICRnu / nu mice),
A carcinostatic agent was previously administered under the same conditions as in Example 2, Evans blue was intravenously administered, and then bradykinin was intradermally administered.
The amount of Evans blue leakage was measured.

(Results) As shown in Table 1, the enhancement of vascular permeability was more remarkable in the normal mice than in the T cell-deficient mice. Furthermore, a VDH response of the auricle was observed in normal mice.

[0048]

[Table 1]

It was found that the action of the host-mediated antitumor agent to promote vascular permeability enhancement of bradykinin is dependent on T cells, as in the expression of antitumor effect by the antitumor agent and VDH reaction.

[0050]

Example 4 The correlation between vascular permeability and VDH response was confirmed using mice having different VDH responses to lentinan.

Similarly to the above, four different strains of mice to which lentinan was administered as an anticancer agent were intravenously administered with Evans blue solution and then bradykinin was subcutaneously administered to examine the degree of vascular hyperpermeability, and at the same time, observe VDH reaction. did.

As a result, as shown in FIG. 3, the vascular responsiveness was different depending on the strain of mouse, and A / J and DB were
In A / 2 mice, a group to which a carcinostatic agent was previously administered (Fig. 3 ●).
In comparison with the non-administered group (Fig. 3 ◯), an increase in vascular permeability due to bradykinin was observed. In contrast, AKR,
In C3H / HeN mice, the effect of pre-administration of an anticancer drug was not observed, and it was confirmed that there was a difference in vascular permeability enhancement depending on the strain of the mouse.

On the other hand, the VDH reaction is A / J and DBA / 2.
C3H / HeN, AK, which was remarkably observed in mice
Not observed in R mice. That is, the increased sensitivity to bradykinin was consistent with the strain difference in VDH reaction.

[0054]

Example 5 A prescription example of the test drug of the present invention containing lentinan will be described. (Prescription) Lentinan 100 μg, Mannitol 20 m
A lyophilized product containing 4 g of dextran and 40.2 mg of dextran is prepared.

(Usage) 0.5 ml of Japanese Pharmacopoeia distilled water for injection is added to the above lyophilized product, and shaken vigorously to dissolve sufficiently. Immediately after the dissolution, 100 μl may be intradermally administered to the subject.

[0056]

Example 6 Next, a prescription example of a test drug containing a vascular permeability-enhancing substance will be described. (Prescription) Bradykinin 5 × 10 ^-9 M, mannitol 20
Prepare a lyophilizate containing mg.

(Usage) 0.5 ml of Japanese Pharmacopoeia distilled water for injection is added to the above lyophilized product, and shaken vigorously to dissolve sufficiently. Immediately after the dissolution, 100 μl may be intradermally administered to the subject.

[0058]

Example 7 Next, a description will be given of an example of prescription of a dye that facilitates measurement of an intradermal reaction. (Prescription) Phenolsulfonephthalein 0.6 g, sodium chloride 0.9 g, sodium hydrogen carbonate 0.143 g
Add distilled water for injection to 100 ml. (Usage) Two minutes after the administration of the test drug, 1.0 ml of the above dye solution is intravenously administered. One hour after the administration, the area of dye-yellow to yellow at the test drug administration site may be measured.

[0059]

[Embodiment 8] Next, a description will be given of an example of prescription of a dye that facilitates the measurement of the intradermal reaction. (Prescription) Distilled water for injection is added to 5 g of sodium fluorescein and 0.9 g of sodium chloride to make 100 ml.

(Usage) Two minutes after the administration of the test agent, the dye solution 2
Administer ml intravenously. One hour after administration, the test drug administration site may be irradiated with ultraviolet rays using a 100 W ultraviolet lamp to measure the fluorescence emission area.

[0061]

INDUSTRIAL APPLICABILITY According to the present invention, the antitumor effect of a host-mediated anticancer drug, which cannot be judged, can be judged at an early stage of treatment with these drugs. As a result, the required dose, the number of doses, the choice of combination method with other drugs,
Alternatively, switching to another drug or the like can be determined, and appropriate treatment can be performed for each patient. This is in line with the policy of medical administration to administer an appropriate drug to an appropriate target patient at an appropriate time.

Further, the test drug of the present invention is excellent in that the test can be carried out quickly without causing any pain to the patient and can be easily carried out in general medical facilities.

[Brief description of drawings]

FIG. 1 is a diagram showing changes in intradermal reaction to administration of lentinan as a test agent by administration of a host-mediated anticancer drug.

FIG. 2 shows changes in sensitivity to bradykinin due to administration of a host-mediated anticancer drug.

FIG. 3 is a diagram showing a mouse strain difference in vascular hyperpermeability.

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Tomoko Kikuchi Tomoko Kikuchi 1-1, Suzuki-cho, Kawasaki-ku, Kanagawa Ajinomoto Co., Inc. Central Research Laboratory (72) Inventor Atsushi Hamuro 1-Suzuki-cho, Kawasaki-ku, Kawasaki-shi, Kanagawa 1 Central Research Institute, Ajinomoto Co., Inc.

Claims (5)

[Claims] 1. An agent for testing an antitumor effect of a host-mediated anticancer agent, which comprises lentinan or sonifilan. 2. The antitumor effect test agent for a host-mediated antitumor agent according to claim 1, wherein the host-mediated antitumor agent is any one of lentinan, sonifilan, picibanil and krestin. 3. An agent for testing an antitumor effect of a host-mediated antitumor agent, which comprises a vascular permeability-enhancing substance. 4. The agent for testing antitumor effect of a host-mediated antitumor agent according to claim 3, wherein the vascular permeability-enhancing substance is any one of bradykinin, kallidin and histamine or a mixture thereof. 5. The agent for testing an antitumor effect of a host-mediated antitumor agent according to claim 3 or 4, wherein the host-mediated antitumor agent is any one of lentinan, sonifilan, picibanil and krestin.