JP3253819B2 - Labeled compound - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a labeled compound and a method for producing the same. More specifically, the present invention relates to a radiochemically labeled compound such as a positron nuclide labeled compound useful for tumor or brain imaging, or a stable isotope labeled compound thereof, a synthetic intermediate thereof, and a production method thereof. .

[0002]

2. Description of the Related Art Conventionally, radioactively labeled substances have been administered to a living body, and the physiological, pharmacological or biochemical processes of trace substances in the living body have been traced in various forms. It has been performed in various ways using compounds. As a method using such a radiochemically labeled compound, recently, a positron-labeled compound is synthesized using a positron (positron) decay (emission) nuclide created by a cyclotron, and administered to the body, followed by positron emission tomography. Imaging equipment (Positron emission tomography, P
In recent years, the PET method for imaging by ET) has attracted attention. Since the positron nuclide is mainly composed of biological constituent elements such as carbon, nitrogen, and oxygen, it is characterized in that various substances and drugs metabolized in the body can be labeled without changing the structure. In addition, due to the nature of emitted annihilation radiation, the above positron tomography apparatus can measure the physiological, pharmacological and biochemical processes of trace substances in the living body with ultra-high sensitivity and extremely low concentration, and can be used clinically. Very useful information can be obtained. One example is the diagnosis of tumors by PET using positron nuclides. It is generally said that tumor cells have higher levels of sugar metabolism, amino acid metabolism, fat metabolism, nucleic acid metabolism, etc. than normal cells, and these metabolisms in tumors directly reflect the goodness of life and the growth status of tumors. Therefore, using a compound in which a sugar or amino acid is labeled with a positron nuclide,
Attempts have been made to diagnose tumors. In such a situation, ¹¹ CL-Methionine is currently commonly used for positron diagnosis of tumors. Recently, 2,4-diaminobutyric acid (DA) is one of the amino acids expected as a specific tumor marker.
BABA), and its uptake into human and rat glioma cells has been observed, and it also has a selective antitumor activity, so it is expected to be effective not only as a tumor marker but also as a new therapeutic agent. One of the amino acids.

However, in practice, the labeled compound used in the PET method has a short half-life of a positron nuclide (for example, 20.4 minutes for ¹¹ C). Although the purity, specific radioactivity, and final radioactivity must be clinically satisfactory, it is actually difficult to produce a positron-labeled compound satisfying such requirements. There is also a problem that workers are exposed to radiation during the synthesis of the labeled compound. For this reason, since a labeling method that is high enough to be practically satisfactory and has a high radiochemical yield has not been established, PET that is expected to be applied to biological observation and diagnosis, for example, imaging of tumors and brains, etc. is expected. The development of the law was not enough.

[0004] For example, its characteristics have attracted attention2,
Positron labeling of 4-diaminobutyric acid (DABA) has also been extremely difficult for these reasons. Regarding the difficulty of such labeling, the situation remains unchanged not only for positron nuclides but also for labeling with radioisotopes such as β-decay nuclides and also stable isotopes.

Accordingly, the present invention solves the above-mentioned problems of the prior art, and provides a new radiolabeled and stable isotope-labeled radioisotope and a stable isotope which can be realized as a labeling method which is rapid and has a high radiochemical yield. It is intended to provide a labeled compound, a synthetic intermediate thereof, and a method for producing them.

[0006]

SUMMARY OF THE INVENTION The present invention, as to solve the above problem, a cyano group carbon ¹¹ C radiochemical target
From known β-cyano-L-alanine or a salt thereof.
The present invention provides a labeled compound characterized by comprising: Further, the present invention provides the following formula (1)

[0007]

Embedded image

(R ₁ is —O-acyl group, —O-alkyl
Group, an amino acid or a salt thereof represented by -O- phosphoryl a group or a halogen atom), a cyano group carbon
¹¹ C radiochemically labeled cyanide and β-cyano-
Are reacted in the presence of L- alanine synthase, preparation of a labeled compound of claim 1, a cyano group carbon is characterized by producing ¹¹ C radiochemical labeled β- cyano -L- alanine or a salt thereof Provide the law.

[0009] Furthermore, the present invention provides the method, wherein the enzyme is a bee.
It is obtained from bacteria belonging to the genus Bacillus
That the bacterium was sent to the Institute of Biotechnology
Bacillus stearothermophilus CN deposited
3 strains (accession number FERM BP-4773)
And the cyanide was ¹¹ C radiochemically labeled
May be selected from hydrocyanic acid, NaCN, or KCN.
As an embodiment of the β-cyano-L-alanine labeled compound,
Offer.

[0010] The present invention also provides any one of the above methods.
Reduce the β-cyano-L-alanine compound produced in
Or the following equation (2)

[0011]

Embedded image

(R ₂ is — ^* CH ₂ —NH ₂ , — ^* CO
NH _2, or - ^* indicates COOH, ^* C is, ¹¹ C radiation
Labeled compounds, characterized in that to produce the amino compound of shows chemical labeled carbon)
Provide a method of manufacturing a product. To explain in more detail,
The labeled β-cyano-L-alanine or
The salt is a novelty provided by this invention for the first time.
A compound represented by the following formula (3)

[0013]

Embedded image

[0014] ^(* C shows the ¹¹ C radiochemical labeled carbon) as represented by the carbon atom of the cyano group, release ¹¹ C
It is characterized by being radiochemically labeled . ¹¹
Β-cyano-L-alanine labeled with C
It is extremely effective as a labeled compound that can be administered in vivo in the PET method produced by conversion of a cyano group. In addition, free amino groups and carboxyl groups are
Naturally, it may be a salt with an acid or an alkali . As described above, the labeled β-cyano-L-alanine as the novel compound can be produced simply and in a high radiochemical yield by the novel production method of the present invention.

That is, in the present invention, for example, an amino acid of the formula (1) or a salt thereof as described above can be prepared by combining a cyano compound having a cyano group carbon labeled with ¹¹ C with β-cyano-L
-It can be produced by reacting in the presence of an alanine synthase. Of course, it goes without saying that it may be produced by so-called chemical synthesis.

In the production of the above labeled β-cyano-L-alanine compound using an enzyme, the enzyme comprises β-cyano-L-alanine.
As those having cyano-L-alanine synthesizing activity, for example, Acinetobacter sp., Acrobacter sp., Agrobacterium sp.
Genus, Arthrobacter, Bacillus, Brevibacterium,
Cellulomonas, Corynebacterium, Enterobacter
Genus, Erwinia, Escherichia, Flavobacterium,
Hafnia, Micrococcus, Mycobacterium, Nocard
Genus ia, Pseudomonas, Rhodococcus, Salmonella
It may be derived from a bacterium selected from the group consisting of the genus, Serratia, and Staphylococcus. Including these, it can also be shown in the following Table 1.

[0017]

[Table 1]

Among them, the enzyme is thermophilic Bacillus (Bac
illus), more specifically, for example, Bacillus stearothermophilus ( Baci
llus Stearothermophilus ). In particular, the bacteria shown as specific examples include Bacillus stearothermophilus strain CN3. The Bacillus stearothermophilus CN3 strain was isolated from nature by the inventor of the present invention,
On March 8, the Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology
Deposited as ERM BP-4773.

Actually, the enzymes for the above reaction obtained from these bacteria include heat-resistant β-
An example is cyanoalanine synthase. 1. Action: Generates β-cyanoalanine from O-acetyl-L-serine and cyanide. Optimum pH: 7.0-9.0 Stable pH: 6.0 to 10.0 4. Optimal temperature: 40-50 ° C 5. Thermal stability: stable up to 70 ° C. when kept at pH 7.5 for 30 minutes Molecular weight: 60,000-80,00 by gel filtration
0.

This enzyme is used to transform a thermophilic bacterium belonging to the genus Bacillus capable of producing β-cyanoalanine synthase into β-cyanoalanine synthase.
-Culturing in a cyanoalanine synthase-inducing medium,
It can be produced by isolating the β-cyanoalanine synthase from the cells. As the bacteria belonging to the thermophilic Bacillus genus in this case, those described above are used, for example. This enzyme requires, for example, pyridoxal phosphate as a coenzyme and O-
Acetyl-L-serine, L-cysteine, L-serine,
O-methyl-L-serine, O-phosphoryl-L-serine, O-succinyl-L-serine or β-chloro-L
-Use alanine or the like as a substrate.

For example, in the reaction of the compound of the formula (1) using the enzyme as described above, the substituent R ₁ of the compound of the formula (1) is more specifically represented by —O-acyl group, —O —An alkyl group, —O-phosphoryl group, or a halogen atom, and the cyan compound can be hydrocyanic acid (CN ⁻ ), NaCN, KCN, or the like, whose carbon is labeled.

In the case of a cyanide labeled with a positron nuclide ¹¹ C as a labeled cyanide, for example, ¹¹ CO ₂ produced by a cyclotron is reduced to ¹¹ CH ₄
And then reacted with ammonia at a high temperature of about 1000 ° C. in the presence of a platinum (Pt) catalyst in the same manner as in a normal hydrocyanic acid synthesis method. This cyanide is reacted with an amino acid of the above formula (1) in an aqueous medium in the presence of an active enzyme as described above, whereby the cyano group carbon as a novel compound of the present invention is a positron nuclide ¹¹ C The β-cyano-L-alanine compound labeled with can be produced. Similarly, a compound labeled with ¹³ C, ¹⁴ C or the like is obtained.

The labeled β-cyano-L-alanine compound is then converted into a compound used in the PET method. In this case, for example, a cyano group can be converted to an amino group by a reduction reaction, or a cyano group can be converted to an amide group or a carboxyl group by a hydrolysis reaction. That is, reduction or decomposition under various conditions produces a labeled amino acid compound of the formula (2), or a salt or a protected derivative thereof by a conventional method.

The reduction reaction can be performed by various means such as a known method such as a method using Raney nickel or Raney cobalt or a method using a reducing agent such as NaBH ₄ . The same applies to the hydrolysis reaction. By these various means including the enzymatic method, for example, conversion into L-2,4-diaminobutyric acid (DABA), γ-aminobutyric acid (GABA), L-asparagine, L-aspartic acid and the like shown below. become.

[0025]

Embedded image

[0026]

According to the present invention, as described above, a cyanide is substituted or added to an amino acid in the presence of an active enzyme, whereby radiochemical or stable isotope labeling of positron nuclide ¹¹ C or the like can be easily performed. And In particular,
The finding that bacteria of the genus Bacillus as a natural microorganism produces enzymes for this reaction is not limited to the positron nuclide ¹¹ C, but also to radioisotopes such as ¹⁴ C and ¹³ C. And labeling with stable isotopes such as

The labeling of various amino acids such as L-2,4-diaminobutyric acid (DABA) greatly contributes not only to observation and diagnosis by the PET method but also to NMR diagnosis and biochemical metabolism research. . I mean,
Conventionally, a method of labeling an amino group or a carboxyl group of an amino acid with an isotope has been known, but these amino groups are easily metabolized in a living body, so that the amino acid mother nucleus could not be traced. . According to the present invention, carbon which is hardly metabolized can be labeled, so that tracking of the mother nucleus can be performed very easily. On the other hand, the mother nucleus beta ^- decay to radioactive isotopes ³ H, or,
It is possible to label for a long time by chemical synthesis with ¹⁴ C or the like, but if these are used, the radioactivity does not penetrate the body, so the position of the labeled compound in the living body is detected from outside the body. It is not possible. ¹¹ C used in the present invention
Since nuclides undergo β ^- decay and collide with the body's negative electrons to emit γ-rays, they can be detected from outside the living body, and therefore, the distribution and localization of the labeled compound administered into the living body can be tracked from outside. It is possible. In fact, for example, the present inventors have administered the obtained ¹¹ CDABA to cultured glioma cells and tumor-bearing rats, and confirmed accumulation in tumor cells. This makes it possible to follow the behavior of the labeled compound in vivo before and after the treatment or without comparing with the clinical effect, which is extremely significant for the diagnosis and treatment of a disease.

Hereinafter, the present invention will be described in more detail with reference to examples.

[0029]

EXAMPLES Reference Example (Heat-resistant β-cyanoalanine synthase) Polypeptone 1
%, Yeast extract 0.25%, glycerol 0.1%,
(NH ₄ ) ₂ SO ₄ 0.1%, MgSO ₄ .7H ₂ O
A medium consisting of 0.05% and K ₂ HPO ₄ 0.1% (pH 7.2) was dispensed into two large test tubes by 8 ml each,
After sterilization at 120 ° C for 20 minutes and cooling, Bacillus stearotherm
One loopful of ophilus CN3 strain (National Institute of Bioscience and Biotechnology, Accession No. FERM BP-4773) was inoculated, and cultured with shaking at 60 ° C. for 24 hours to obtain a seed culture solution. 1.6 L of a medium obtained by adding 0.01% (V / V) of an antifoaming agent (Adecanol KG-126) to the same medium composition as above was placed in a 2 L jar fermenter, and
After sterilization and cooling at 20 ° C. for 20 minutes, the seed culture solution 1 was added thereto.
6 ml (for 2 test tubes) was inoculated, 60 ° C, 27 hours,
Culture was performed under the conditions of an aeration rate of 1.6 L / min and a stirring speed of 300 rpm to obtain a pre-culture liquid. Next, polypeptone 1%, yeast extract 0.25%, glycerol 0.1%, (N
H ₄ ) ₂ SO ₄ 0.1%, MgSO ₄ .7H ₂ O 0.0
5%, K ₂ HPO ₄ 0.1%, L-serine 0.1% (p
160 L of the medium consisting of H7.2) was placed in a 200-L jar fermenter, sterilized at 120 ° C. for 30 minutes, cooled, and inoculated with 1.6 L of the above pre-cultured solution. Culture was performed under the conditions of aeration and a stirring speed of 200 rpm. After the completion of the culture, the cells were collected by centrifugation using a sharp press.

The obtained cells were divided into eight equal portions (20 L each), and an appropriate amount of a potassium phosphate buffer solution (10 m
M, pH 8.0, containing 0.1 mM dithiothreitol)
And stored frozen at -80 ° C. Unzip this,
It was used for the subsequent production of the enzyme. After suspending 60 L of the frozen cells to a total volume of about 2000 ml, the cells were disrupted by Dyno-mill. The crushed liquid was centrifuged to remove bacterial cell residues to obtain 2100 ml of a cell-free extract.

The cell-free extract was added with 4 parts of ammonium sulfate.
0% saturation was added, and after standing overnight, the precipitated precipitate was removed by centrifugation. Ammonium sulfate was again added to the obtained supernatant to 90% saturation, left for 5 hours, and centrifuged. A precipitate was obtained. The precipitate is added to 0.1
It was dissolved in a 10 mM potassium phosphate buffer (pH 8.0) containing mM dithiothreitol, and desalted with a buffer of the same composition using a dialysis membrane. Final concentration 70 in 1065 ml of desalted solution
% Ethanol was previously cooled to -80 ° C, and a precipitate was obtained by centrifugation. This is 0.1
The suspension was suspended in 10 mM potassium phosphate buffer (pH 8.0) containing mM dithiothreitol, and heat-treated at 70 ° C. for 30 minutes. After removing the precipitate by centrifugation, the supernatant 1024
ml was passed through a DEAE-Cellulofine A500 column (6.0 cm in diameter x 18 cm in length) which had been equilibrated with 10 mM potassium phosphate buffer (pH 8.0) containing 0.1 mM dithiothreitol in advance to adsorb the enzyme. After washing with a buffer of the composition, 0.1 mM dithiothreitol and 0.5 M NaCl were added from a 10 mM potassium phosphate buffer (pH 8.0) containing 0.1 mM dithiothreitol.
100 mM potassium phosphate buffer (pH 8.0) containing
The enzyme was eluted by a gradient elution method, and the active fraction was collected.

To the active fraction was added 80% ammonium sulfate.
The mixture was added so as to be saturated, and after standing overnight, a precipitate was obtained by centrifugation. This precipitate was dissolved in 10 mM potassium phosphate buffer (pH 8.0) containing 0.1 mM dithiothreitol, and subjected to preparative electrophoresis (7.5% polyacrylamide gel). After the electrophoresis, an active portion in the gel was cut out, crushed, and the enzyme was extracted using a 10 mM potassium phosphate buffer (pH 8.0) containing 0.1 mM dithiothreitol. Ammonium sulfate was added to this active fraction so as to become 30% saturated, and phenyl sepharose CL-4B previously equilibrated with 10 mM potassium phosphate buffer (pH 8.0) containing 30% saturated ammonium sulfate and 0.1 mM dithiothreitol. Column (diameter 2.
5 cm x 12 cm) to adsorb the enzyme, wash with a buffer of the same composition, and remove from a 10 mM potassium phosphate buffer (pH 8.0) containing 30% saturated ammonium sulfate and 0.1 mM dithiothreitol. 10 mM potassium phosphate buffer containing 1 mM dithiothreitol (pH
The enzyme was eluted by the gradient elution method described in (8.0), and the active fraction was collected.

[0033] Ammonium sulfate was added to this active fraction at 30%.
10m containing 30% saturated ammonium sulfate and 0.1 mM dithiothreitol in advance
Octyl Sepharose CL-4B column (1.5 cm diameter) equilibrated with M potassium phosphate buffer (pH 8.0)
× length 10 cm) to adsorb the enzyme, wash with a buffer of the same composition, and add 30% saturated ammonium sulfate and 0.1 m
Enzyme was eluted by a gradient elution method from 10 mM potassium phosphate buffer (pH 8.0) containing M dithiothreitol to 10 mM potassium phosphate buffer (pH 8.0) containing 0.1 mM dithiothreitol. Fractions were collected. It was confirmed that the obtained enzyme preparation was single electrophoretically.

Table 2 shows the enzyme activity, yield, and the like for each step in the above process of obtaining the enzyme. Further, “Unit” in Table 2 is described in Table 3
According to the activity measurement method, the enzyme activity for producing 1 μmol of β-cyanoalanine per minute was defined as one unit (Unit).

[0035]

[Table 2]

[0036]

[Table 3]

The obtained enzyme is O-acetyl-L
-It has the properties shown in the following Table 4 as having β-cyano-L-alanine synthesizing activity from serine and a cyano compound.

[0038]

[Table 4]

[0039]Example 1 Positron nuclides produced by cyclotron¹¹Hold C
One¹¹CO_TwoAnd H_TwoAnd N_TwoN under mixed atmosphere
reducing at 400 ° C. in the presence of i.¹¹CH _FourConversion to
And at a temperature of 1000 ° C., platinum (Pt)
Contact reaction with ammonia as a catalyst, H¹¹Manufacture CN
did. Mixed gaseous H¹¹50% H_TwoSO_Foursolution
1.5 ml to remove residual ammonia,
Further P _TwoO_FiveTo remove ammonia
And add H to 350 μl of 50 mM KOH.¹¹Trap CN
Was.

O-acetyl-L-serine was mixed with the aqueous solution of H ¹¹ CN together with the β-cyano-L-alanine synthesizing enzyme obtained in the above Reference Example and reacted at room temperature. The product was analyzed under the conditions in Table 5 below.

[0041]

[Table 5]

FIGS. 1 and 2 show the results of analysis at UV 220 nm and Radiodetector. This figure 1
2 and FIG. 2 confirmed the presence of a β-cyano-L-alanine structure and a cyano group having a positron nuclide ¹¹ C. FIG. 3 shows a mass spectrometry spectrum. In addition to the results of FIGS. 1 and 2, FIG. 3 also shows that β-cyano-L in which the cyano group carbon was labeled with a positron nuclide ¹¹ C.
-It was confirmed to be alanine. Example 2 The carbon of the cyano group obtained in Example 1 is a positron nuclide
Β-cyano-L-alanine labeled with ¹¹ C has C
Reduction treatment was performed by adding oBr ₂ and NaBH ₄ reducing agents. Next, after filtration (0.2 μm), 500 μl of 6 M hydrochloric acid was added, and the mixture was filtered through a 0.2 μm filter to collect the enzyme, which was purified by HPLC.

The product was analyzed under the conditions shown in Table 6 below.

[0044]

[Table 6]

FIGS. 4 and 5 show the results of analysis using UV at 220 nm and a Radiodetector. From this result, it was confirmed that it was L-2,4-diaminobutyric acid (L-DABA) labeled with ¹¹ C. This compound had a radiochemical purity of 96% or more and a radiochemical yield of 30 to 40%. FIG. 6 shows the identification of L-2,4-diaminobutyric acid (L-DABA) and D-2,4-diaminobutyric acid (D-DABA).
And the values analyzed by Radiodetector.

2,4-enzymatically synthesized according to the present invention
Diaminobutyric acid (DABA) has been proven to be the L-form. The chart on the left in FIG.
Derivatization of 4-diaminobutyric acid (DABA) and HPLC
It is what saw. The method for derivatizing the D, L-form is described in Ma.
rfey P. Carlsberg Res. Commun. 49,591 (1984).

The right chart in FIG. 6 shows that the enzymatically synthesized labeled DABA is L-form DABA. About ¹¹ C-labeled L-2,4-diaminobutyric acid obtained by Example 3 Example 2 (L-DABA), were evaluated for biological applicability as labeling substance. For rat glioma, medium with amino acid content close to that of living body and Bu for comparison
The relationship between the concentration and radioactivity of ffer phosphate buffered saline (PBS) was examined. FIG. 7 shows the result.

The above-mentioned Medi of ¹¹ CL-DABA of the present invention
From the results of um, it can be seen that ¹¹ CL-DABA has good properties as a labeling substance applicable to living organisms.

[0049]

As described above in detail, according to the present invention, it is possible to produce an amino acid compound such as L-2,4-diaminobutyric acid labeled with positron nuclide ¹¹ C or the like, which has been difficult to produce until now. It is possible simply and with a high radiochemical yield. In addition, a labeled β-cyano-L-alanine compound, which is a synthetic intermediate, is provided to facilitate the production of the amino acid compound.

It facilitates observation of in vivo behavior and greatly contributes to disease diagnosis and treatment.

[Brief description of the drawings]

FIG. 1. UV of ¹¹ C-labeled β-cyano-L-alanine
It is a spectrum diagram.

FIG. 2. Radiation of ¹¹ C-labeled β-cyano-L-alanine
It is an odetector spectrum figure.

FIG. 3 is a mass spectrometry diagram of ¹¹ C-labeled β-cyano-L-alanine.

FIG. 4. UV of ¹¹ C-labeled L-2,4-diaminobutyric acid
It is a spectrum diagram.

FIG. 5. Radiation of ¹¹ C-labeled L-2,4-diaminobutyric acid
It is a spectrum diagram of odetector.

FIG. 6 is a spectrum diagram of UV-340 nm and Radiodetector of L-2,4-diaminobutyric acid and D-2,4-diaminobutyric acid.

FIG. 7: glio of ¹¹ C-labeled L-2,4-diaminobutyric acid
FIG. 4 is a concentration-radioactivity correlation diagram showing the nature of application in ma.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI C12P 13/06 (C12N 9/88 // (C12N 9/88 C12R 1:07) C12R 1:07) A61K 49/02 Z ( 72) Inventor Keigo Komura 2-179, Minamimatsunaga-cho, Fukuyama-shi, Hiroshima 2 (72) Inventor Yuji Furuya 20-9 Jusangenya, Kanbe-cho, Shenzhen-gun, Hiroshima 20-72 (72) Inventor Bengt Longström Sweden Uppsala S -751 85 Inside the Uppsala University PT Center (72) Inventor Gunner Antoni Sweden Uppsala S-751 85 Inside the Uppsala University PT Center (56) References JP-A-54-100316 (JP, A) Label. Compounds, Vol. 5, No. 2 (1969), p. 142-151J. Labeled Compd. Radiopharm. , Vol. 22, No. 9 (1985), p. 909-915 Med. Chem. , Vol. 25 (1982), p. 93-96 (58) Field surveyed (Int. Cl. ⁷ , DB name) A61K 49/00-49/04 CA (STN) REGISTRY (STN)

Claims (6)

(57) [Claims] 1. The method according to claim 1, wherein the carbon of the cyano group is chemically labeled with ¹¹ C.
consisting of β-cyano-L-alanine or a salt thereof
Characteristic labeled compounds . 2. The following formula (1) : (R ₁ represents —O-acyl group, —O-alkyl group, —O—
A phosphoryl group or a halogen atom) or a salt thereof,
¹¹ C radiochemically labeled cyanide and β-cyano-
By reacting in the presence of L-alanine synthase, a cyano group
Β-cyano-L-alani carbon radiochemically labeled with ¹¹ C
Or a salt thereof is produced.
A method for producing a labeled compound . 3. The enzyme belongs to the genus Bacillus.
Claims characterized by being obtained from bacteria
2. Manufacturing method . 4. The method according to claim 1, wherein said bacteria are
Bacillus stearothermophy deposited at the institute
Lus CN3 strain (Accession number FERM BP-4773)
The method of claim 3 . 5. The method according to claim 5, wherein the cyanide is an ¹¹ C radiochemically labeled.
Selected from hydrocyanic acid, NaCN, or KCN
The method according to any one of claims 2 to 4, wherein: 6. The method according to claim 2, wherein
Reducing or decomposing the compound of claim 1 to form
(2) ## STR2 ## (R _{2 ^{is, - * CH 2 -NH 2,}} - * CONH 2, also
The - ^* indicates COOH, ^* C is ¹¹ C radiochemical labeling
Labeled compounds, characterized in that to produce the amino compound of indicating carbon)
Manufacturing method of goods .