JPS6320839B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to the synthesis of novel N-benzyl and N,N-dibenzyl derivatives of adriamycin and daunomycin and to compositions containing said derivatives and pharmaceutically acceptable acid addition salts, which derivatives are useful as anti-cancer chemicals. It is utilized and has the following structure. In the formula, R is _-COCH3 , -CHOH- _CH3 , -
It represents COCH ₂ OH or -CHOH-CH ₂ OH, and X represents -NHCH ₂ Ph or -N(CH ₂ Ph) ₂ (herein "Ph" represents a phenyl group). For ease of identification, eight compounds within the scope of the present invention are listed in Table 1 below.

Table The preparation of the compounds is described in detail in the Examples. However, it is noted here that the reductive alkylation of daunomycin () with benzaldehyde in the presence of sodium cyanoborohydride for an extended reaction period at room temperature
The objective is to produce good yields of the N,N-dibenzyl compound () and the 13-dihydro derivative () as a by-product. For relatively short reaction periods,
Mono-N-benzyldaunomycin and mono-N-benzyldaunomycin predominate. By selecting the stoichiometric amount, either N-benzylation or N,N-dibenzylation can be obtained. mono-N-benzylation,
N,N-dibenzyladriamycin () by treating adriamycin () with benzylaldehyde and sodium cyanoborohydride under conditions that predominantly yield compound ().
was similarly obtained. Reductive alkylation is achieved by ketone reduction, resulting in four products (
) were observed in the benzylation of daunomycin, and four additional species (,
,,) were observed from the benzylation of adriamycin. Although identified by analysis,
No compound was isolated. However, it is believed to have anti-cancer activity comparable to that of other compounds of the invention. The compounds of the invention may be prepared in either the free base or acid addition salt form. The salts are soluble in water and, for example, aqueous propylene glycol, while the free base form of the compound is soluble in selected organic solvents such as chloroform, methylene chloride and ethyl acetate. The salts are thus particularly well suited for the treatment of animals, including humans, since they can be used in the form of aqueous solutions, including salines. These acid addition salts (herein prepared as acid addition salts of HCl) are preferably pharmaceutically acceptable non-toxic addition salts with suitable acids, such as inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid. , sulfuric acid and phosphoric acid and organic acids such as organic carboxylic acids such as glycolic acid, maleic acid, hydroxymaleic acid, malic acid, tartaric acid, citric acid, salicylic acid, and organic sulfonic acids such as methanesulfonic acid and toluene-p-sulfonic acid. salts. Acid addition salts may be converted into the free compounds according to known methods, for example by converting metal hydroxides or alkoxides such as alkali metal or alkaline earth metal hydroxides, such as lithium hydroxide, sodium hydroxide, water bases such as potassium oxide or calcium hydroxide; alkali metal or alkaline earth metal carbonates or acidic carbonates such as sodium, potassium, or calcium carbonates or acidic carbonates; ammonia; or hydroxyl ion exchange resins, or other suitable and treating the acid addition salt with a reagent. Acid addition salts may also be converted into other acid addition salts according to known methods, e.g. salts with inorganic acids are It can be treated with metal salts such as sodium, barium or silver salts. Acid addition salts can be converted into other acid addition salts by treatment with anion exchange processes. The preparation of each of the above compounds is described in the following examples. Example 1 N-benzyldaunomycin (), N-benzyl-13-dihydrodaunomycin () and N,N-dibenzyldaunomycin (), and HCl salts. Daunomycin/HCl (2.26 g, 4.0 mmol)
and benzaldehyde (8.2 mL, 80.0 mmol)
was taken up in 3:1 acetonitrile/H ₂ O (116 mL) and stirred at 23° C. for 0.5 h.
NaCNBH ₃ (0.75 g, 12.0 mmol) was added and stirring continued for 0.5 h. the reactant
Dilute with _H2O (150 mL) and CHCl3 ( ₃ x 100
mL). The extracts were combined and added to _H2O
(30 mL) and the aqueous phase was washed with CHCl ₃ (50 mL).
mL). The organic phases were combined and extracted with cold 0.1N HOAc (6 x 100 mL). The extracts were combined, immediately made basic with NaHCO ₃ and extracted with CHCl ₃ (3×300 mL). The extracts were combined, evaporated and the residue chromatographed (1.3Kg silica gel - 50mm x
7ft dry column - 10:1 _CHCl3 /MeOH), 1.06g N-benzyldaunomycin () (51%) and
0.17 g of N-benzyl-13-dihydrodaunomycin (7%) was obtained as free base.
The organic solution after HOAc extraction was dried and evaporated. The residue was chromatographed (560 g silica gel - 50 mm x 3 ft dry column - 10:1 CHCl ₃ /
MeOH), 0.18 g (6%) of N,N-dibenzyldaunomycin () free base was obtained. The HCl salts were prepared by reacting a CHCl ₃ solution of each compound with an equivalent amount of methanolic HCl followed by precipitation with ether. 1.28g (49%) ();
[α] _D +252° ( c 0.05, 95% EtOH); tlc (10:
1CHCl ₃ /MeOH)R _f 0.60, Analysis Calculated value of C ₃₄ H ₃₅ NO ₁₀・HCl・H ₂ O: C H N Cl 60.76 5.70 2.08 5.28 Measured value: 60.88 5.59 1.99 5.40 0.16g (6%) of () ; [α] _D +207゜( c 0.047
,
95% EtOH); tlc (10:1 CHCl ₃ /MeOH) R _f 0.4, Analysis Calculated value of C ₃₄ H ₃₇ NO ₁₀・HCl・1.25H ₂ O: C H N Cl 60.17 6.02 2.06 5.22 Measured value: 60.07 5.79 2.26 5.38 0.16g (5%) (); [α] _D +283゜( c 0.048
,
95% EtOH); tlc (10:1 CHCl ₃ /MeOH) R _f 0.90, Analysis Calculated value of C ₄₁ H ₄₁ NO ₁₀・HCl・0.75H ₂ O: C H N Cl 64.99 5.79 1.85 4.68 Measured value: 65.08 5.69 1.77 4.45 Example 2 N-Benzyl Adriamycin (), N,N
-dibenzyladriamycin (), and N
-benzyl-13-dihydroadriamycin () and HCl salts. Adriamycin HCl (2.90g, 50mmol)
and benzaldehyde (10.5 mL, 100 mmol)
was taken up in 2:1 acetonitrile/H ₂ O (150 mL) and stirred for 20 minutes at 23°C.
NaCNBH ₃ ( _0.94 g,
15.0 mmol) and benzaldehyde (10.5 mmol)
L) was added to the solution. Stirring was continued for 30 minutes and the reaction was diluted with H ₂ O (150 mL) and extracted with CHCl ₃ (4×100 mL).
The extracts were combined and saturated NaCl (2 x 100 mL)
Washed with water, dried and evaporated. The residue was chromatographed (1.3Kg silica gel - 50mm
×7ft dry column - 40:10: _1CHCl3 /MeOH/
H ₂ O), 0.10 g (3%) N,N-dibenzyladriamycin (), 1.02 g (30%) N-benzyladriamycin (), and 1.20 g (38%)
N-benzyl-13-dihydroadriamycin () was obtained as the free base. The elution after () was a small amount of material that was not completely purified but was tentatively identified as N,N-dibenzyl-13-dihydroadriamycin (). These were treated with HCl by the treatment described in the previous example.
Converted to salt yielding: 0.07g (2%) (); [α] _D +275゜( c 0.051
,
85% EtOH); tlc (10:1 CHCl ₃ /MeOH) R _f 0.85, Analysis Calculated value of C ₄₁ H ₄₁ NO ₁₁・HCl・0.6H ₂ O: C H N Cl 63.86 5.64 1.81 4.59 Measured value: 63.88 5.57 1.83 4.45 0.90g (28%) (); [α] _D +209゜( c 0.048
,
95% EtOH); tlc (10:1 CHCl ₃ /MeOH) R _f 0.55, Analysis Calculated value of C ₃₄ H ₃₅ NO ₁₁・HCl・H ₂ O: C H N Cl 59.34 5.56 2.03 5.15 Measured value: 59.46 5.40 2.01 5.37 1.00g (30%) (); [α] _D +204゜( c 0.050
,
95% EtOH); tlc (10:1 CHCl ₃ /MeOH) R _f 0.25, Analysis Calculated value of C ₃₄ H ₃₇ NO ₁₁・HCl・0.8H ₂ O: C H N Cl 59.48 5.87 2.04 5.16 Measured value: 59.43 5.81 2.05 5.40 Example 3 N,N-dibenzyldaunomycin (), N,
N-dibenzyl-13-dihydrodaunomycin (), N-benzyldaunomycin (), and N-benzyl-13-dihydrodaunomycin (), and HCl salts. Daunomycin HCl (2.54 g, 4.5 mmol)
and benzaldehyde (9.2 mL, 90.0 mmol)
was taken up in 2:1 acetonitrile/H ₂ O (90 mL) and stirred for 20 minutes at 23°C. NaCNBH ₃ (0.57 g, 9.0 mmol) in acetonitrile (20 mL) was added dropwise over 20 minutes.
was added to the dissolved stirred solution and stirring was continued for 14 days. The reaction was dissolved in H ₂ O (150 mL).
and extracted with CHCl ₃ (5 x 100 mL). The extracts were combined, washed with water (100 mL), and the aqueous phase was extracted with CHCl ₃ (50 mL). The organic solutions were combined, dried and evaporated and the residue was chromatographed (190 g silica gel - 2.8 x 64 cm column - CHCl ₃ to 9:
_1CHCl3 /MeOH), 1.19g (38%) (),
0.41 g (13%) of N,N-dibenzyl-13-dihydrodaunomycin (), 0.30 g (11%) of () and 0.10 g (4%) of () were obtained as free base in the order of their elution. . These were converted to HCl salts by the procedure described above to yield: 1.20 g (35%) () and 0.37 g (11%) (); [α] _D + 247° ( c 0.049, 95% EtOH); tlc
(10:1CHCl ₃ /MeOH) R _f 0.85, analysis Calculated value of C ₄₁ H ₄₃ NO ₁₀・HCl・1.5H ₂ O: C H N Cl 63.68 6.13 1.81 4.58 Measured value: 63.66 5.75 1.78 4.86 and 0.23 as HCl salt g (8%) () and 0.07 g (2%) (). The compounds of the invention, including their salts, can be administered to animals by any available route, including oral and parenteral (intravenous, intraperitoneal, subcutaneous and intramuscular) administration. The dose is an amount sufficient to cure leukemia or other types of cancer in which the compound may be found to be effective, and the dose is dependent on the type of cancer, the type of animal, and the weight of the animal. Depends on. For example, in human administration, approximately 0.1
A dose of a compound of the invention within the range of mg/Kg to about 500 mg/Kg is sufficient to ameliorate leukemia. A similar dosage range is therapeutically effective in treating lower test animals. The upper dosage limit is imposed by toxic side effects and can be determined by trial and error on the animal to be treated, including humans. To facilitate administration, the compounds of the invention, including their salts, may be presented in the form of compositions and preferably in dosage units. Although any of the selected compounds can be administered by itself, it is commonly administered with a pharmaceutically acceptable carrier that dilutes the compound and facilitates handling. The term "pharmaceutically acceptable" means that the carrier (as well as the resulting composition) is sterile and non-toxic. The carrier or diluent may be solid, semi-solid, or liquid and may serve as a vehicle, excipient form, or medium for the anticancer agent. Examples of diluents and carriers include lactose, dextrose, sucrose, sorbit, mannitol, starches, gum acacia, calcium phosphate, mineral oil, cocoa butter, cocoa butter, Mention may be made of alginates, tragacanth, gelatin, syrup, methylcellulose, polyoxyethylene sorbitan, monolaurate, methyl- and propyl-hydroxybenzoate, talc and magnesium stearate. For convenient handling, particularly when intended for use in dosage units, the compound and carrier or diluent may be packaged in a capsule, sachet, cachet, gelatin, paper or other container. Can be encapsulated. The dosage unit may take the form of a tablet, capsule, suppository or cachet, for example. The following examples illustrate various forms of dosage units. Here, the HCl salt of the compound may be formulated. Example 4 Tablet Formulation Mg/Tablet Compound 15 Lactose 86 Cornstarch (dried) 45.5 Gelatin 2.5 Magnesium Stearate 1.0 The compound was powdered and passed through a mesh sieve.
and both sieved lactose and 30
Mix thoroughly with mg cornstarch. The above mixed powder is lumped with a hot gelatin solution prepared by stirring and heating the gelatin in water to make a 10% w/w solution. The mass is granulated by passing through a sieve and the wet granules are dried at 40°C. Regranulate the dried granules by passing through a sieve, add the starch and the remainder of the magnesium stearate, and mix thoroughly. The granules are compressed to form tablets each weighing 150 mg. Example 5 Tablet Formulation Mg/Tablet Compound 100 Lactose 39 Cornstarch (dried)
80 Gelatin 4.0 Magnesium Stearate 2.0 The formulation is the same as that of Example 4, except that 60 mg of starch is used in the granulation process and 20 mg during tabletting. Example 6 Capsule Formulation Mg/Capsule Compound 250 Lactose 150 The compound and lactose are passed through a sieve and mixed well together before filling into hard gelatin capsules of appropriate size and each capsule is
Contain 400mg of mixed powder. Example 7 Suppositories Mg/Suppositories Compound 50 Cocoa Butter 950 The compound was powdered, passed through a sieve, and 45
C. to form an evenly mixed suspension by grinding with molten cocoa butter. The mixture was stirred well and each nominally 1
Make suppositories by pouring into g capacity molds. Example 8 Cassier Mg/Casier Compound 100 Lactose 400 The compound is passed through a mesh sieve, mixed with the lactose previously passed through the sieve, and placed into appropriately sized Cassiers each containing 500 mg. Example 9

【table】 Example 10

Other compounds of the invention could be formulated in dosage unit form in the same general manner as described above for the compounds. Biological Testing All samples except compound () were prepared for biological screening and recent test results, both in vitro and in vivo, are shown in Table 2 below. It can be seen that each compound is active against leukemia P388 in rats and that N,N-dibenzyldaunomycin (compound) is clearly the best. What appears to be surprising and highly significant in the in vitro test results is that when proceeding from N-benzyl to N,N-dibenzyl compounds,
There is a sudden and unexpected loss of DNA binding. Although ΔT _n is the most direct measure of DNA binding, inhibition of nucleic acid synthesis is also lost. Most of the anthracyclines are likely to have an effect on DNA by an intercalation mechanism.
binds to and causes mutation. Although these N,N-dibenzyl compounds retain their antitumor activity in mouse selection,
DNA binding is lost. This suggests that N,N-dibenzyl compounds may be useful and effective anticancer agents, but may act by a different mechanism than most of the other anthracyclines, and thus Serious cardiotoxicity common to all
This means that side effects can be avoided. The compound described herein as daunomycin () is otherwise known as daunorubicin or rubidomycin.

【table】

Claims (1)

[Claims] 1. The following structure [Wherein, R is _-COCH3 , -CHOH- _CH3 , -
COCH ₂ OH or -CHOH-CH ₂ OH, and X represents -NHCH ₂ Ph or -N(CH ₂ Ph) ₂ ] or a pharmaceutically acceptable acid addition salt thereof. 2. The compound according to claim 1, or a pharmaceutically acceptable acid addition salt thereof, which is N-benzyldaunomycin or a pharmaceutically acceptable acid addition salt thereof. 3. The compound according to claim 1, or a pharmaceutically acceptable acid addition salt thereof, which is N-benzyl-13-dihydrodaunomycin or a pharmaceutically acceptable acid addition salt thereof. 4 N-Benzyl Adriamycin or a pharmaceutically acceptable acid addition salt thereof, the compound according to claim 1 or a pharmaceutically acceptable acid addition salt thereof. 5. The compound according to claim 1 or a pharmaceutically acceptable acid addition salt thereof, which is N-benzyl-13-dihydroadriamycin or a pharmaceutically acceptable acid addition salt thereof. 6 N,N-dibenzyldaunomycin or a pharmaceutically acceptable acid addition salt thereof, Claim 1
Compounds listed in Section 1 or their pharmaceutically acceptable acid addition salts. 7. The compound according to claim 1, or a pharmaceutically acceptable acid addition salt thereof, which is N,N-dibenzyl-13-dihydrodaunomycin or a pharmaceutically acceptable acid addition salt thereof. 8. The compound according to claim 1 or a pharmaceutically acceptable acid addition salt thereof, which is N,N-dibenzyladriamycin or a pharmaceutically acceptable acid addition salt thereof. 9. The compound according to claim 1, or a pharmaceutically acceptable acid addition salt thereof, which is N,N-dibenzyl-13-dihydroadriamycin or a pharmaceutically acceptable acid addition salt thereof. 10 dosage units of a pharmaceutical composition for treating leukemia, the composition comprising:
a therapeutically effective amount of at least one compound to ameliorate leukemia in the range of 0.1 to about 500 mg, together with a pharmaceutically acceptable non-toxic carrier or diluent, wherein the at least one compound is N-benzyl. Daunomycin, N-benzyl-13-dihydrodaunomycin, N-benzyladriamycin, N-benzyl-13-dihydroadriamycin, N,N-dibenzyldaunomycin, N,
N-dibenzyl-13-dihydrodaunomycin,
N,N-dibenzyladriamycin and N,N
- A pharmaceutical composition selected from the group consisting of dibenzyl-13-dihydroadriamycin and their pharmaceutically acceptable acid addition salts. 11. A pharmaceutical composition according to claim 10, wherein the dosage unit is in the form of a tablet. 12. A pharmaceutical composition according to claim 10, wherein the dosage unit is in the form of a capsule. 13. A pharmaceutical composition according to claim 10, wherein the dosage unit is in the form of a suppository. 14. A pharmaceutical composition according to claim 10, wherein the dosage unit is in the form of a cachet. 15. The pharmaceutical composition of claim 10, wherein the dosage unit is in sterile aqueous form. 16 Claim 15 in the form of an aqueous solution
Pharmaceutical compositions as described in Section. 17 Claim 15 in aqueous dispersion form
Pharmaceutical compositions as described in Section.