RU2799339C2 - Prodrugs of opioid receptor antagonists - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions includes prodrugs of opioid receptor antagonists of the formula:

,

where X represents O or CH₂ or their pharmaceutically acceptable salts. The group of inventions also relates to a pharmaceutical composition containing the said prodrugs, a method for treating opioid dependence, and the use of the pharmaceutical composition.

EFFECT: provision of opioid receptor antagonists that provide long-term, delayed and controlled release of nalmefene and naltrexone opioid receptor antagonists when administered.

14 cl, 101 tbl, 49 dwg, 73 ex

Description

BACKGROUND OF THE INVENTION

[0001] There is a need in the medical field for compositions and techniques to modulate the activity of opioid receptors in the treatment of behavioral disorders.

SUMMARY OF THE INVENTION

[0002] The present invention provides prodrugs of opioid receptor antagonists such as nalmefene and naltrexone, pharmaceutical compositions containing said compounds, and methods of using said compounds to treat behavioral disorders.

[0003] Some compounds of the present invention have better characteristics. For example, some compounds of the present invention are more stable in pharmaceutical compositions based on oils such as sesame oil or cottonseed oil.

[0004] Some compounds of the present invention have more favorable pharmacokinetic characteristics in vivo (eg, in rats or dogs), such as a prolonged half-life.

[0005] Some compounds of the present invention have better in vivo safety parameters (eg, in rats or dogs), such as a less pronounced reaction at the injection site.

[0006] Certain compounds of the present invention are more stable, either alone or in oil-based pharmaceutical compositions.

[0007] According to one aspect of the present invention, there is provided a compound, or a pharmaceutically acceptable salt thereof, having the structure represented by Formula (II),

(II),

(II)

Where

X is O or CH ₂ ;

,R represents

,

where R ¹ is C ₄ -C ₁₀ alkyl or C ₄ -C ₁₀ alkenyl and n is 7-15, provided that if X is O then n is not 7.

[0008] In some embodiments, X is O. In some embodiments, X is CH ₂ .

[0009] According to another aspect of the present invention, there is also provided a compound, or a pharmaceutically acceptable salt thereof, having the structure represented by Formula (IIa),

(IIa),

(IIa),

Where

X is O or CH ₂ ;

R represents

,

,

where R ¹ is C ₄ -C ₁₀ alkyl or C ₄ -C ₁₀ alkenyl and n is 9-15.

[0010] In some embodiments, X is O. In some embodiments, X is CH ₂ .

[0011] According to another aspect of the present invention, there is also provided a compound, or a pharmaceutically acceptable salt thereof, having the structure represented by Formula (III),

(III),

(III)

Where

X is O or CH ₂ ;

R is selected from

-[CH(R ³ )O]zR ⁴ ;

-[CH(R ³ )O]zC(=O)OR ⁴ ;

-[CH(R ³ )O]zC(=O)NR ⁴ R ⁵ and

-[CH(R ³ )O]zP(=O)(OR ⁴ )(OR ⁵ ),

where z is 1, 2, 3, 4, 5, 6, or 7;

R ³ represents a hydrogen atom, a halogen atom, alkyl, alkenyl, cycloalkylalkyl or aryl;

each R ⁴ and R ⁵ is independently selected from hydrogen, alkyl, alkenyl, cycloalkylalkyl or aryl.

[0012] In some embodiments, X is O. In some embodiments, X is CH ₂ .

[0013] According to one embodiment, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound represented by any of Formulas (II), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these connections.

[0014] The present invention provides a method for treating opioid dependence in a patient in need of such treatment, comprising administering a pharmaceutical composition containing a compound of Formula (II), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt said compounds, and a pharmaceutically acceptable excipient.

[0015] In one embodiment, the present invention provides a method of treating a patient, wherein the therapeutic effect of the deposited long acting opioid antagonist can be abolished in the patient by administration of an opioid analgesic.

[0016] According to one embodiment, the present invention provides a method for treating opioid dependence in a patient in need of such treatment, characterized in that the patient receives a first injection of an injectable preparation containing a compound represented by any of Formulas (II), (IIa) or (III ), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these compounds, providing maintenance of a therapeutically significant plasma concentration for about 1 week, about 2 weeks, about 3 weeks, or about 4 weeks, at least about 2 months, according to at least about 3 months, at least about 4 months, at least about 5 months, or at least about 6 months, and then a second injection of an injectable preparation containing a compound represented by any of Formulas (II), (IIa) or ( III), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these compounds, providing maintenance of a therapeutically significant plasma concentration for at least about 2 months, at least about 3 months, at least about 4 months, at least at least about 5 months, or at least about 6 months.

[0017] According to one embodiment, the present invention provides a method for treating opioid dependence in a patient in need of such treatment, characterized in that the patient receives a first injection of an injectable preparation of naltrexone enclosed in PLGA microspheres, which maintains a therapeutically relevant plasma concentration for approximately 4 weeks, and then one or more injections of an injectable preparation containing a compound represented by any of Formulas (II), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these compounds, providing maintaining a therapeutically relevant plasma concentration for about 2 months, about 3 months, about 4 months, or about 5 months or more.

[0018] According to one embodiment, the present invention provides a method for treating opioid dependence in a patient in need of such treatment, characterized in that the patient receives one or more injections of an injectable preparation containing at least one compound represented by any of Formulas (II ), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these compounds, and the patient has previously received treatment with opioid agonists or partial agonists such as buprenorphine or methadone, and the patient is currently in transition to discontinuation of treatment with such agonists or partial agonists.

[0019] According to one embodiment, the present invention provides a method for treating opioid dependence in a patient in need of such treatment, characterized in that the patient receives one or more injections of an injectable preparation containing at least one compound represented by any of Formulas (II ), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these compounds, and the patient has recently become dependent and has not previously received medication in combination with behavioral therapy, or the patient has recently stopped taking opioid pain medication, is at risk of future opioid drug dependence and needs to prevent future opioid drug dependence by treatment with antagonists.

inclusion by reference

[0020] All publications, patents and patent applications mentioned in this description are incorporated by reference into this description for the specific purposes indicated in this description.

BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Various aspects of the present invention are detailed in the appended claims. A better understanding of the advantages and features of the present invention may be obtained from the following detailed description, which sets forth illustrative embodiments of the invention using the principles of the invention, and the accompanying drawings. The patent application set contains at least one color drawing. Copies of this patent application publication with color drawing(s) may be obtained from the office upon request, at a fee.

[0022] In FIG. 1 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl ((E)-octadec-9-en-1-yl) carbonate (Example 1).

[0023] In FIG. 2 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl undecyl carbonate (Example 2).

[0024] In FIG. 3 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl undecyl carbonate (Example 3).

[0025] In FIG. 4 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl undecyl carbonate (Example 4).

[0026] In FIG. 5 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl undecyl carbonate (Example 5).

[0027] In FIG. 6 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl dodecyl carbonate (Example 6).

[0028] In FIG. 7 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl ((E)-octadec-9-en-1-yl) carbonate (Example 7).

[0029] In FIG. 8 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl (E)-octadec-9-enoate (Example 8).

[0030] In FIG. 9 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl (E)-octadec-9-enoate (Example 9).

[0031] In FIG. 10 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl decyl carbonate (Example 10).

[0032] In FIG. 11 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl dodecyl carbonate (Example 11).

[0033] In FIG. 12 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl stearate (Example 12).

[0034] In FIG. 13 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl (Z)-docos-13-enoate (Example 13).

[0035] In FIG. 14 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl docosanoate (Example 14).

[0036] In FIG. 15 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl (E)-octadec-9-enoate (Example 15).

[0037] In FIG. 16 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl eicosanoate (Example 16).

[0038] In FIG. 17 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl octyl carbonate (Example 17).

[0039] In FIG. 18 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl decyl carbonate (Example 18).

[0040] In FIG. 19 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl hexadecyl carbonate (Example 19).

[0041] In FIG. 20 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl (9Z,12Z,15Z)-octadeca-9,12,15-trienoate (Example 20).

[0042] In FIG. 21 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl hexadecyl carbonate (Example 21).

[0043] In FIG. 22 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl (Z)-docos-13-enoate (Example 22).

[0044] In FIG. 23 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl octyl carbonate (Example 23).

[0045] In FIG. 24 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecyl carbonate (Example 24).

[0046] In FIG. 25 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl tetradecyl carbonate (Example 25).

[0047] In FIG. 26 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl (E)-octadec-9-enoate (Example 26).

[0048] In FIG. 27 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl tetradecyl carbonate (Example 27).

[0049] In FIG. 28 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl eicosyl carbonate (Example 28).

[0050] In FIG. 29 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecyl carbonate (Example 29).

[0051] In FIG. 30 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl tridecyl carbonate (Example 30).

[0052] In FIG. 31 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl tetradecyl carbonate (Example 31).

[0053] In FIG. 32 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl pentadecyl carbonate (Example 32).

[0054] In FIG. 33 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl octadecyl carbonate (Example 33).

[0055] In FIG. 34 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecyl carbonate (Example 34).

[0056] In FIG. 35 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl decyl carbonate (Example 35).

[0057] In FIG. 36 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl oleate (Example 36).

[0058] In FIG. 37 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl (9Z,12Z)-octadeca-9,12-dienoate (Example 37).

[0059] In FIG. 38 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3,3-dimethylbutanoate (Example 38).

[0060] In FIG. 39 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3-cyclopentylpropanoate (Example 39).

[0061] In FIG. 40 shows the nuclear magnetic resonance spectrum of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl tert-butyl carbamate (Example 40).

[0062] In FIG. 41 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl oleate (Example 41).

[0063] In FIG. 42 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3,3-dimethylbutanoate (Example 42).

[0064] In FIG. 43 shows the nuclear magnetic resonance spectrum of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4, 12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3-cyclopentylpropanoate (Example 43).

[0065] In FIG. 44 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecanoate (Example 44).

[0066] In FIG. 45 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl tetradecanoate (Example 45).

[0067] In FIG. 46 shows the nuclear magnetic resonance spectrum (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecanoate (Example 46).

[0068] In FIG. 47 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecyl carbonate (Example 47).

[0069] In FIG. 48 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecanoate (Example 48).

[0070] In FIG. 49 shows the nuclear magnetic resonance spectrum (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H- 4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecanoate (Example 49).

DETAILED DESCRIPTION OF THE INVENTION

[0071] In the context of this description and the appended claims, the singular forms of nouns include the corresponding plural forms, unless the meaning clearly implies otherwise. Accordingly, reference to, for example, the word "agent" implies that a plurality of such agents are included, and reference to the word "cell" implies that one or more cells (or a plurality of cells) and equivalent formulations known to those skilled in the art are included, and so Further. When ranges of physical properties such as molecular weight or chemical properties such as chemical formulas are used throughout this specification, all combinations and subcombinations of ranges and ranges found in specific embodiments of the invention are intended to be included. The term "approximately" referring to a number or range of numerical values means that the specified number or range of numerical values is an approximation within the experimental variability (or within the statistical experimental error), and, accordingly, the number or range of numerical values, in some cases , may differ by 1-15% from the specified number or range of numeric values. The term "comprising" (and related terms such as "comprise" or "comprises" or "having" or "comprising") is not intended to exclude that in some other embodiments of the invention, for example in an embodiment of any composition substances, composition, method or process, or the like, described in this application, "consist of" the described components or "consist essentially of" the described components.

definitions

[0072] In the context of this description and the attached claims, unless otherwise indicated, the following terms have the following meanings.

[0073] "Amino" refers to the radical-NH ₂ .

[0074] "Cyano" refers to the -CN radical.

[0075] "Nitro" refers to the -NO ₂ radical.

[0076] "Oxa" refers to the -O- radical.

[0077] "Oxo" refers to the radical =O.

[0078] "Thioxo" refers to the =S radical.

[0079] "Imino" refers to the radical =N-H.

[0080] "Oximo" refers to the radical =N-OH.

[0081] "Hydrazino" refers to the radical =N-NH ₂ .

[0082] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, containing no unsaturations, having from one to fifteen carbon atoms (eg C ₁ -C ₁₅ alkyl). In some embodiments, alkyl contains from one to thirteen carbon atoms (eg C ₁ -C ₁₃ alkyl). In some embodiments, alkyl contains from one to eight carbon atoms (eg C ₁ -C ₈ alkyl). According to other embodiments of the invention, alkyl contains from one to five carbon atoms (eg C ₁ -C ₅ alkyl). According to other embodiments of the invention, alkyl contains from one to four carbon atoms (eg C ₁ -C ₄ alkyl). According to other embodiments of the invention, alkyl contains from one to three carbon atoms (eg C ₁ -C ₃ alkyl). According to other embodiments of the invention, alkyl contains from one to two carbon atoms (eg C ₁ -C ₂ alkyl). According to other embodiments of the invention, alkyl contains one carbon atom (eg C ₁ alkyl). According to other embodiments of the invention, alkyl contains from five to fifteen carbon atoms (eg C ₅ -C ₁₅ alkyl). According to other embodiments of the invention, alkyl contains from five to eight carbon atoms (eg C ₅ -C ₈ alkyl). According to other embodiments of the invention, alkyl contains from two to five carbon atoms (eg C ₂ -C ₅ alkyl). According to other embodiments of the invention, alkyl contains from three to five carbon atoms (eg C ₃ -C ₅ alkyl). According to other embodiments of the invention, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2 -methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl is connected to the rest of the molecule through a single bond. Unless otherwise noted herein, an alkyl group optionally has one or more of the following substituents: halogen, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR ^a , -SR ^a , -C (O) -R ^a , -N(R ^a ) ₂ , -C(O)R ^a , -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -N(R ^a )C(O) OR ^a , - C (O)-N(R ^a ) ₂ , -N(R ^a )C(O)R ^a , -N(R ^a )S(O) _t R ^a (where t is 1 or 2 ), -S(O) _t OR ^a (where t is 1 or 2), -S(O) _t R ^a (where t is 1 or 2), and -S(O) _t N(R ^a ) ₂ (where t is 1 or 2) wherein each R ^a is independently hydrogen, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with a halogen atom, hydroxy , methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom , hydroxy, methoxy, or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen, hydroxy, methoxy, or trifluoromethyl atom).

[0083] "Alkoxy" refers to an oxygen-linked radical represented by Formula -O-alkyl, where alkyl is an alkyl chain as defined above.

[0084] "Alkenyl" refers to a radical group having a straight or branched hydrocarbon chain, consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and having from two to twelve carbon atoms. In some embodiments, the alkenyl contains two to eight carbon atoms. According to other embodiments of the invention, alkenyl contains from two to four carbon atoms. Alkenyl is connected to the rest of the molecule via a single bond, examples are ethenyl (i.e. vinyl), prop-1-enyl (i.e. allyl), but-1-enyl, penta-1-enyl, penta-1,4-dienyl and the like. Unless otherwise noted herein, an alkenyl group optionally has one or more of the following substituents: halogen, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR ^a , -SR ^a , - C (O) -R ^a , -N(R ^a ) ₂ , -C(O)R ^a , -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -N(R ^a )C(O) OR ^a , - C (O)-N(R ^a ) ₂ , -N(R ^a )C(O)R ^a , -N(R ^a )S(O) _t R ^a (where t is 1 or 2 ), -S(O) _t OR ^a (where t is 1 or 2), -S(O) _t R ^a (where t is 1 or 2), and -S(O) _t N(R ^a ) ₂ (where t is 1 or 2) wherein each R ^a is independently hydrogen, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with a halogen atom, hydroxy , methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom , hydroxy, methoxy, or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen, hydroxy, methoxy, or trifluoromethyl atom).

[0085] "Alkynyl" refers to a radical group having a straight or branched hydrocarbon chain, consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond having from two to twelve carbon atoms. In some embodiments, the alkynyl contains two to eight carbon atoms. In other embodiments, the alkynyl contains two to six carbon atoms. In other embodiments, the alkynyl contains two to four carbon atoms. The alkynyl is connected to the rest of the molecule via a single bond, examples are ethynyl, propynyl, butynyl, pentynyl, hexynyl and the like. Unless otherwise noted herein, the alkynyl group optionally has one or more of the following substituents: halogen, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR ^a , -SR ^a , - C (O) -R ^a , -N(R ^a ) ₂ , -C(O)R ^a , -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -N(R ^a )C(O) OR ^a , - C (O)-N(R ^a ) ₂ , -N(R ^a )C(O)R ^a , -N(R ^a )S(O) _t R ^a (where t is 1 or 2 ), -S(O) _t OR ^a (where t is 1 or 2), -S(O) _t R ^a (where t is 1 or 2), and -S(O) _t N(R ^a ) ₂ (where t is 1 or 2) wherein each R ^a is independently hydrogen, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with a halogen atom, hydroxy , methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom , hydroxy, methoxy, or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen, hydroxy, methoxy, or trifluoromethyl atom).

[0086] "Alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain linking the rest of the molecule and a radical group consisting solely of carbon and hydrogen atoms, containing no unsaturations and having from one to twelve carbon atoms, examples are methylene, ethylene, propylene, n-butylene and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The attachment of the alkylene chain to the rest of the molecule and to the radical group is carried out through one carbon atom of the alkylene chain or through any two carbon atoms of this chain. In some embodiments, the alkylene contains from one to eight carbon atoms (eg C ₁ -C ₈ alkylene). According to other embodiments of the invention, the alkylene contains from one to five carbon atoms (eg C ₁ -C ₅ alkylene). According to other embodiments of the invention, the alkylene contains from one to four carbon atoms (eg C ₁ -C ₄ alkylene). According to other embodiments of the invention, the alkylene contains from one to three carbon atoms (eg C ₁ -C ₃ alkylene). According to other embodiments of the invention, the alkylene contains from one to two carbon atoms (eg C ₁ -C ₂ alkylene). According to other embodiments of the invention, the alkylene contains one carbon atom (eg C ₁ alkylene). In other embodiments, the alkylene contains five to eight carbon atoms (eg C ₅ -C ₈ alkylene). According to other embodiments of the invention, the alkylene contains from two to five carbon atoms (eg C ₂ -C ₅ alkylene). According to other embodiments of the invention, the alkylene contains from three to five carbon atoms (eg C ₃ -C ₅ alkylene). Unless otherwise noted herein, the alkylene chain optionally contains one or more of the following substituents: halogen, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR ^a , -SR ^a , - C (O) -R ^a , -N(R ^a ) ₂ , -C(O)R ^a , -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -N(R ^a )C(O) OR ^a , - C (O)-N(R ^a ) ₂ , -N(R ^a )C(O)R ^a , -N(R ^a )S(O) _t R ^a (where t is 1 or 2 ), -S(O) _t OR ^a (where t is 1 or 2), -S(O) _t R ^a (where t is 1 or 2), and -S(O) _t N(R ^a ) ₂ (where t is 1 or 2) wherein each R ^a is independently hydrogen, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with a halogen atom, hydroxy , methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom , hydroxy, methoxy, or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen, hydroxy, methoxy, or trifluoromethyl atom).

[0087] "Alkynylene" or "alkynylene chain" refers to a straight or branched divalent hydrocarbon chain linking the remainder of the molecule and a radical group consisting solely of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond and having one up to twelve carbon atoms. The alkynylene chain is attached to the rest of the molecule via a single bond and to the radical group via a single bond. In some embodiments, the alkynylene contains two to eight carbon atoms (eg C ₂ -C ₈ alkynylene). In other embodiments, the alkynylene contains two to five carbon atoms (eg C ₂ -C ₅ alkynylene). In other embodiments, the alkynylene contains two to four carbon atoms (eg C ₂ -C ₄ alkynylene). In other embodiments, the alkynylene contains two to three carbon atoms (eg C ₂ -C ₃ alkynylene). According to other embodiments of the invention alkynylene contains two carbon atoms (eg C ₂ alkylene). In other embodiments, the alkynylene contains five to eight carbon atoms (eg C ₅ -C ₈ alkynylene). In other embodiments, the alkynylene contains three to five carbon atoms (eg C ₃ -C ₅ alkynylene). Unless otherwise stated in this specification, the alkynylene chain optionally contains one or more of the following substituents: halogen, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl, -OR ^a , -SR ^a , - C (O) -R ^a , -N(R ^a ) ₂ , -C(O)R ^a , -C(O)OR ^a , -C(O)N(R ^a ) ₂ , -N(R ^a )C(O) OR ^a , - C (O)-N(R ^a ) ₂ , -N(R ^a )C(O)R ^a , -N(R ^a )S(O) _t R ^a (where t is 1 or 2 ), -S(O) _t OR ^a (where t is 1 or 2), -S(O) _t R ^a (where t is 1 or 2), and -S(O) _t N(R ^a ) ₂ (where t is 1 or 2) wherein each R ^a is independently hydrogen, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted with a halogen atom, hydroxy , methoxy or trifluoromethyl), carbocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom , hydroxy, methoxy, or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen, hydroxy, methoxy, or trifluoromethyl atom).

[0088] "Aryl" refers to a radical resulting from an aromatic monocyclic or polycyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. This aromatic monocyclic or polycyclic hydrocarbon ring system contains only hydrogen and carbon atoms, with the number of carbon atoms ranging from five to eighteen, and at least one of the rings in said ring system is completely unsaturated, i.e. contains a cyclic, delocalized (4n+2 ) π-electron system in accordance with Hückel's theory. The ring system from which the aryl groups are derived includes, without limitation, groups such as benzene, fluorene, indane, indene, tetralin, and naphthalene. Unless otherwise specified herein, the term "aryl" or the prefix "ar-" (e.g., in the term "aralkyl") is intended to include aryl radicals optionally containing one or more substituents independently selected from alkyl, alkenyl, alkynyl, halogen atom, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkynyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroaryl alkyl , -R ^b -OR ^a , -R ^b - C (O) -R ^a , -R ^b - C (O) -OR ^a , -R ^b - C (O) -N(R ^a ) ₂ , -R ^b -N(R ^a ) ₂ , -R ^b -C(O)R ^a , -R ^b -C(O)OR ^a , -R ^b -C(O)N(R ^a ) ₂ , -R ^b - OR ^c -C(O)N(R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , -R ^b - N(R ^a )S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O ) _t OR ^a (where t is 1 or 2) and -R ^b -S(O) _t N(R ^a ) ₂ (where t is 1 or 2), where each R ^a independently represents a hydrogen atom, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), cycloalkylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl ), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), each R ^b is independently a straight bond or a straight or branched alkylene or alkenylene chain and R ^c is a straight or branched alkylene or alkenylene chain, and each of the above substituents is unsubstituted unless otherwise indicated.

[0089] "Aralkyl" refers to a radical represented by the Formula -R ^c -aryl, where R ^c represents an alkylene chain as defined above, for example, methylene, ethylene, and the like. The alkylene chain portion of the aralkyl radical is optionally substituted in accordance with the above definition of an alkylene chain. The aryl moiety of an aralkyl radical is optionally substituted in accordance with the above definition of an aryl group.

[0090] "Aralkenyl" refers to a radical represented by the Formula -R ^d -aryl, where R ^d is an alkenylene chain as defined above. The aryl moiety of the aralkenyl radical is optionally substituted in accordance with the above definition of an aryl group. The alkenylene chain portion of the aralkenyl radical is optionally substituted in accordance with the above definition of an alkenylene group.

[0091] "Aralkynil" refers to a radical represented by Formula -R ^e -aryl, where R ^e is an alkynylene chain as defined above. The aryl moiety of the aralkynyl radical is optionally substituted according to the above definition of an aryl group. The alkynylene chain portion of the aralkynyl radical is optionally substituted in accordance with the above definition of an alkynylene chain.

[0092] "Aralkoxy" refers to an oxygen-linked radical represented by the Formula -OR ^c -aryl, where R ^c is an alkylene chain such as defined above, eg, methylene, ethylene, and the like. The alkylene chain portion of the aralkyl radical is optionally substituted in accordance with the above definition of an alkylene chain. The aryl moiety of an aralkyl radical is optionally substituted in accordance with the above definition of an aryl group.

[0093] "Carbocyclyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical consisting solely of carbon and hydrogen atoms, which includes fused or bridged ring systems, having from three to fifteen carbon atoms. In some embodiments, the carbocyclyl contains from three to ten carbon atoms. In other embodiments, the carbocyclyl contains five to seven carbon atoms. Carbocyclyl is connected to the rest of the molecule through a single bond. Carbocyclyl is saturated (ie contains only single CC bonds) or unsaturated (ie contains one or more double bonds or triple bonds). A fully saturated carbocyclyl radical is also referred to as "cycloalkyl". Examples of monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Unsaturated carbocyclyl is also referred to as "cycloalkenyl". Examples of monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. Polycyclic carbocyclyl radicals include, for example, adamantyl, norbornyl (ie, bicyclo[2.2.1]heptanyl), norbornenyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise specified herein, the term "carbocyclyl" is intended to include carbocyclyl radicals which optionally contain one or more substituents independently selected from alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkynyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R ^b -OR ^a , - R ^b - C (O) -R ^a , -R ^b - C (O) -OR ^a , -R ^b - C (O) -N (R ^a ) ₂ , -R ^b -N (R ^a ) ₂ , -R ^b -C(O)R ^a , -R ^b -C(O)OR ^a , -R ^b -C(O)N(R ^a ) ₂ , -R ^b -OR ^c -C(O)N( R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , -R ^b -N(R ^a )S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O) _t OR ^a (where t is 1 or 2) and -R ^b -S(O) _t N(R ^a ) ₂ (where t is 1 or 2), where each R ^a independently represents a hydrogen atom, alkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with substituents halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), each R ^b independently represents is a direct bond or a straight or branched alkylene or alkenylene chain; and R ^c is a straight or branched alkylene or alkenylene chain, and each of the above substituents is unsubstituted unless otherwise indicated.

[0094] "Carbocyclylalkyl" refers to a radical represented by the Formula -R ^c -carbocyclyl, where R ^c is an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical are optionally substituted according to the above definitions.

[0095] "Carbocyclylalkynyl" refers to a radical represented by Formula -R ^c -carbocyclyl, where R ^c is an alkynylene chain as defined above. The alkynylene chain and the carbocyclyl radical are optionally substituted according to the above definitions.

[0096] "Carbocyclylalkoxy" refers to an oxygen-linked radical represented by the Formula -OR ^c -carbocyclyl, where R ^c is an alkylene chain as defined above. The alkylene chain and the carbocyclyl radical are optionally substituted according to the above definitions.

[0097] As used herein, the term "carboxylic bioisostere" refers to a functional group or moiety that has physical, biological and/or chemical properties similar to those of a carboxyl moiety. Examples of carboxyl bioisosteres include, without limitation,

и тому подобное.

etc.

[0098] The term "halogen" or "halogen atom" refers to bromine, chlorine, fluorine or iodine substituents.

[0099] "Fluoroalkyl" refers to an alkyl radical as defined above which is substituted with one or more fluoro radicals as defined above, examples are trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2 -trifluoroethyl, 1-fluoromethyl-2-fluoroethyl and the like. According to some embodiments of the invention, the alkyl portion of the fluoroalkyl radical is optionally substituted in accordance with the above definition of an alkyl group.

[00100] "Heterocyclyl" refers to a stable 3-18 membered non-aromatic ring radical that contains two to twelve carbon atoms and one to six heteroatoms selected from nitrogen, oxygen and sulfur atoms. Unless otherwise specified herein, a heterocyclyl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which optionally includes fused or bridged ring systems. The heteroatoms of the heterocyclyl radical are optionally oxidized. One or more nitrogen atom, if present, is optionally quaternized. The heterocyclyl radical is partially or fully saturated. The heterocyclyl is connected to the rest of the molecule through any atom of the ring/rings. Examples of such heterocyclyl radicals include, without limitation, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrroles dinyl, oxazolidinyl, piperidinyl , piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specified herein, the term "heterocyclyl" is meant to include heterocyclyl radicals as defined above, which optionally contain one or more substituents selected from alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, oxo -R ^b -OR ^a , -R ^b -°C (O)-R ^a , -R ^b -°C (O) -OR ^a , -R ^b -°C (O)-N(R ^a ) ₂ , -R ^b -N(R ^a ) ₂ , -R ^b -C(O)R ^a , -R ^b -C(O)OR ^a , -R ^b -C(O)N(R ^a ) ₂ , -R ^b -OR ^c -C(O)N(R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , -R ^b -N (R ^a )S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O) _t OR ^a (where t is 1 or 2) and -R ^b -S(O) _t N(R ^a ) ₂ (where t is 1 or 2), where each R ^a independently represents a hydrogen atom, alkyl ( optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl) , aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl ), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), each R ^b is independently a straight bond or a straight or branched alkylene or alkenylene chain; and R ^c is a straight or branched alkylene or alkenylene chain, and each of the above substituents is unsubstituted unless otherwise indicated.

[00101] "N-heterocyclyl" or "N-attached heterocyclyl" refers to a heterocyclyl radical, as defined above, containing at least one nitrogen atom and characterized in that the attachment of the heterocyclyl radical to the rest of the molecule is through the nitrogen atom of the heterocyclyl radical. The N-heterocyclyl radical is optionally substituted according to the above definition of heterocyclyl radicals. Examples of such N-heterocyclyl radicals include, without limitation, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

[00102] "C-heterocyclyl" or "C-attached heterocyclyl" refers to a heterocyclyl radical, as defined above, containing at least one heteroatom and characterized in that the attachment of the heterocyclyl radical to the rest of the molecule is through the carbon atom of the heterocyclyl radical . C-heterocyclyl radical is optionally substituted in accordance with the above definition of heterocyclyl radicals. Examples of such C-heterocyclyl radicals include, without limitation, 2-morpholinyl, 2- or 3- or 4-piperidinyl, 2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

[00103] "Heterocyclylalkyl" refers to a radical represented by the Formula -R ^c -heterocyclyl, where R ^c is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, then the site of attachment of this heterocyclyl to the alkyl radical is optionally located at the nitrogen atom. The alkylene chain of the heterocyclylalkyl radical is optionally substituted in accordance with the above definition of an alkylene chain. The heterocyclyl portion of the heterocyclylalkyl radical is optionally substituted according to the above definition of a heterocyclyl group.

[00104] "Heterocyclylalkoxy" refers to an oxygen-linked radical represented by the Formula -OR ^c -heterocyclyl, where R ^c is an alkylene chain as defined above. If the heterocyclyl is a nitrogen-containing heterocyclyl, then the site of attachment of this heterocyclyl to the alkyl radical is optionally located at the nitrogen atom. The alkylene chain of the heterocyclylalkoxy radical is optionally substituted in accordance with the above definition of an alkylene chain. The heterocyclyl moiety of the heterocyclylalkoxy radical is optionally substituted in accordance with the above definition of a heterocyclyl group.

[00105] "Heteroaryl" refers to a radical derived from a 3-18 membered aromatic ring radical that contains two to seventeen carbon atoms and one to six heteroatoms selected from nitrogen, oxygen and sulfur atoms. In the context of this description, a heteroaryl radical is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, where at least one of the rings of the ring system is completely unsaturated, that is, it contains a cyclic, delocalized (4n+2) π-electron system in accordance with the theory Hückel. Heteroaryl includes fused or bridged ring systems. The heteroatom(s) of the heteroaryl radical are optionally oxidized. One or more nitrogen atom, if present, is optionally quaternized. The heteroaryl is connected to the rest of the molecule through any atom of the ring/rings. Examples of heteroaryls include, without limitation, azepinyl, acridinyl, benzimidazolyl, benzdolyl, 1,3-benzodioxolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1 ,4]oxazinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxynyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6] imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[ h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3, 2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8, 9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranil, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7, 8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl and thiophenyl (i.e. thienyl). Unless otherwise specified herein, the term "heteroaryl" is meant to include heteroaryl radicals as defined above, which optionally contain one or more substituents selected from alkyl, alkenyl, alkynyl, halogen, fluoroalkyl, haloalkenyl , haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkynyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroa rilalkyl , -R ^b -OR ^a , -R ^b -°C (O) -R ^a , -R ^b -°C (O) -OR ^a , -R ^b -°C (O) -N(R ^a ) ₂ , -R ^b -N(R ^a ) ₂ , -R ^b -C(O)R ^a , -R ^b -C(O)OR ^a , -R ^b -C(O)N(R ^a ) ₂ , - R ^b -OR ^c -C(O)N(R ^a ) ₂ , -R ^b -N(R ^a )C(O)OR ^a , -R ^b -N(R ^a )C(O)R ^a , - R ^b -N(R ^a )S(O) _t R ^a (where t is 1 or 2), -R ^b -S(O) _t R ^a (where t is 1 or 2), -R ^b - S(O) _t OR ^a (where t is 1 or 2) and -R ^b -S(O) _t N(R ^a ) ₂ (where t is 1 or 2), where each R ^a is independently an atom hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with a halogen atom, hydroxy , methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with a halogen atom, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with a halogen atom, hydroxy, methoxy, or trifluoromethyl), each R ^b is independently a straight bond or a straight or branched alkylene or alkenylene chain and R ^c is a straight or branched alkylene or alkenylene chain, and each of the above substituents is unsubstituted unless otherwise indicated.

[00106] "N-heteroaryl" refers to a heteroaryl radical, as defined above, containing at least one nitrogen atom and characterized in that the attachment of the heteroaryl radical to the rest of the molecule is through the nitrogen atom of the heteroaryl radical. The N-heteroaryl radical is optionally substituted according to the above definition of heteroaryl radicals.

[00107] "C-heteroaryl" refers to a heteroaryl radical, as defined above, characterized in that the attachment of the heteroaryl radical to the rest of the molecule is through the carbon atom of the heteroaryl radical. C-heteroaryl radical is optionally substituted in accordance with the above definition of heteroaryl radicals.

[00108] "Heteroarylalkyl" refers to a radical represented by the Formula -R ^c -heteroaryl, where R ^c is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, then the site of attachment of this heteroaryl to the alkyl radical is optionally located at the nitrogen atom. The alkylene chain of the heteroarylalkyl radical is optionally substituted in accordance with the above definition of an alkylene chain. The heteroaryl moiety of the heteroarylalkyl radical is optionally substituted according to the above definition of a heteroaryl group.

[00109] "Heteroarylalkoxy" refers to an oxygen-linked radical represented by the Formula -OR ^c -heteroaryl, where R ^c is an alkylene chain as defined above. If the heteroaryl is a nitrogen-containing heteroaryl, then the site of attachment of this heteroaryl to the alkyl radical is optionally located at the nitrogen atom. The alkylene chain of the heteroarylalkoxy radical is optionally substituted in accordance with the above definition of an alkylene chain. The heteroaryl moiety of the heteroaryl alkoxy radical is optionally substituted in accordance with the above definition of a heteroaryl group.

[00110] The compounds described in this application, according to some embodiments of the present invention, contain one or more asymmetric center and, accordingly, can exist as enantiomers, diastereoisomers and other stereoisomeric forms, the absolute configuration of which can be defined in terms of stereochemistry as ( R)- or (S)-. Unless otherwise indicated, the present invention is intended to include all stereoisomeric forms of the compounds described in this application. When the compounds described in this application contain alkene double bonds, and unless otherwise indicated, this invention is meant to include both E and Z geometric isomers (eg cis or trans). Moreover, it is intended that all possible isomers as well as their racemic and optically pure forms and all tautomeric forms are also included. The term "geometric isomer" refers to the E or Z geometric isomers (eg cis or trans) relative to the alkene double bond. The term "positional isomer" refers to structural isomers with respect to the central ring, such as ortho, meta and para isomers with respect to the benzene ring.

[00111] The term "tautomer" refers to a molecule in which the transfer of a proton from one atom of the molecule to another atom of the same molecule is possible. The compounds described herein, according to some embodiments of the invention, exist as tautomers. In those cases where tautomerization is possible, a chemical equilibrium of the tautomers is usually established. The exact ratio of tautomers depends on several factors including physical state, temperature, solvent, and pH. Some examples of tautomeric equilibrium include:

.
,

,

,

,

,

,

.

[00112] The compounds described herein are used in various enriched isotopic forms in some embodiments of the invention, for example, compounds enriched in ² H isotopes are used. In one specific embodiment of the present invention, the compound is deuterated at at least one position. Such deuterated forms can be prepared using the methodology described in US Pat. Nos. 5,846,514 and 6,334,997. As described in US Pat.

[00113] According to another embodiment of the present invention, the compounds described in this application have some or all of ^{the 1} H atoms replaced by ² H atoms. Methods for the synthesis of deuterium-containing compounds are known in the art and include the following synthetic procedures, given as a non-limiting Example.

[00114] Deuterium-substituted compounds are synthesized using various techniques, such as those described in the following publications: Dean, Dennis C. (Editor), Recent Advances in the Synthesis and Applications of Radiolabeled Compounds for Drug Discovery and Development [in Curr. Pharm. Des., 2000; 6(10)] 2000, 110 pp.; George W., Rajender S. Varma, The Synthesis of Radiolabeled Compounds via Organometallic Intermediates, Tetrahedron, 1989, 45(21), 6601-21; and Evans E. Anthony, Synthesis of radiolabeled compounds, J. Radioanal. Chem., 1981, 64(1-2), 9-32.

[00115] Deuterated starting materials are readily available and can be used in the synthetic procedures described herein to synthesize deuterium-containing compounds. A large number of deuterium-containing reagents and building blocks are commercially available from chemical suppliers such as Aldrich Chemical Co.

[00116] Deuterium transfer reagents suitable for use in nucleophilic displacement reactions, such as iodomethane-d ₃ (CD ₃ I), are readily available and can be used to transfer a deuterium-substituted carbon atom to a reaction substrate under the conditions of nucleophilic displacement reactions. The use of CD ₃ I is shown below in the Reaction Schemes given by way of example only.

base

base

[00117] Deuterium transfer reagents such as lithium aluminum deuteride (LiAlD ₄ ) are used to transfer deuterium to the reaction substrate under reducing conditions. The use of LiAlD ₄ is shown below in the Reaction Schemes, given by way of example only.

[00118] Deuterium gas and a palladium catalyst are used to reduce unsaturated carbon-carbon bonds and perform reductive substitution of aryl carbon-halogen bonds, as shown below in the Reaction Schemes, given by way of example only.

[00119] According to one embodiment of the invention, the compounds described in this application contain one atom of deuterium. According to another embodiment of the invention, the compounds described in this application contain two deuterium atoms. According to another embodiment of the invention, the compounds described in this application contain three deuterium atoms. According to another embodiment of the invention, the compounds described in this application contain four deuterium atoms. According to another embodiment of the invention, the compounds described in this application contain five deuterium atoms. According to another embodiment of the invention, the compounds described in this application contain six deuterium atoms. According to another embodiment of the invention, the compounds described in this application contain more than six deuterium atoms. According to another embodiment of the invention, the compounds described in this application are completely deuterium-substituted and do not contain non-exchangeable ¹ H hydrogen atoms. According to one embodiment of the invention, the level of incorporation of deuterium is determined using synthetic procedures that use a deuterated synthetic building block as a starting material.

[00120] A "pharmaceutically acceptable salt" includes acid and base addition salts. A pharmaceutically acceptable salt of any of the compounds described herein that are prodrugs of opioid receptor antagonists is intended to include any and all pharmaceutically acceptable salt forms. Preferred pharmaceutically acceptable salts of the compounds described in this application are pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[00121] The term "pharmaceutically acceptable acid addition salt" refers to those salts that retain the biological efficacy and properties of free bases, are not biologically or otherwise undesirable, and are formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid, hydrofluoric acid, phosphoric acid, and the like. Also included are salts formed with organic acids such as aliphatic monobasic and dibasic carboxylic acids, phenyl-substituted monobasic carboxylic acids, monobasic hydroxycarboxylic acids, dibasic carboxylic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and so on, which include salts formed , e.g. with acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. Representative salts respectively include sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates, monohydrogen phosphates, dihydrogen phosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, trifluoroacetates, propionates, caprylates, isobutyrates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, mandelates, benzoates, chlorobenzoates, methyl benzoates, dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates, phenylacetates, citrates, lactates, malates, tartrates, methanesulfonates, and the like. Also included are amino acid salts such as arginates, gluconates, and galacturonates (see, for example, Berge S. M. et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 66:1-19 (1997)). Acid addition salts of basic compounds, in some embodiments of the invention, are prepared according to methods and procedures familiar to those skilled in the art, including contacting the compound in free base form with a sufficient amount of the desired acid to form said salt.

[00122] The term "pharmaceutically acceptable base addition salt" refers to those salts that retain the biological effectiveness and properties of free acids and are not biologically or otherwise undesirable. These salts are obtained by adding an inorganic base or an organic base to a free acid. Pharmaceutically acceptable base addition salts, according to some embodiments of the present invention, are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Salts derived from inorganic bases include, without limitation, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, e.g., salts of isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine , 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, N,N-dibenzylethylenediamine, chlorprocaine, hydrabamine, choline, betaine, ethylenediamine, ethylenedianiline, N-methylglucamine, glucosamine, methylglucamine, theobromine, purines , piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. See above Berge et al.

[00123] In the context of this description, the terms "treatment" or "treat" or "temporary relief" or "attenuation" are used interchangeably. These terms refer to a technique for obtaining beneficial or desired results, including, without limitation, a therapeutic effect and/or a prophylactic effect. "Therapeutic effect" means the elimination or amelioration of the causative disorder being treated. Also, a therapeutic effect is considered achieved if one or more physiological symptoms associated with a given causative disorder are eliminated or reduced, so that the patient experiences an improvement in condition, despite the fact that the patient still has not eliminated the causative disorder itself. To achieve a prophylactic effect, the compositions, according to some embodiments of the present invention, are administered to a patient at risk of developing a particular disease, or to a patient who has one or more physiological symptoms of the disease, even if the disease has not yet been diagnosed.

pharmacology of opioid receptors

[00124] The opioid receptors, μ, δ, κ and the opioid-like receptor ORL-1, belong to the superfamily of G protein coupled receptors (GPCRs) that have seven helical transmembrane domains in their architecture. Most of the research focusing on this group of proteins has focused on the μ receptor, as it mediates the action of both the opiate and opioid analgesics such as morphine and fentanyl, respectively. However, over time, it became more and more obvious that all members of this family of proteins are actively involved in a huge number of biological processes. In addition, the emergence of selective antagonists has demonstrated that both negative and positive modulation of this receptor family provides pharmacotherapeutic possibilities.

[00125] The μ receptor (mu, OP ₃ or MOP) was first identified and characterized pharmacologically by Martin, Kosterlitz et al. based on the high affinity and sensitivity of this receptor for morphine (Martin et al. The effects of morphine- and nalorphine-like drugs in the independent and morphine-dependent chronic spinal dog (J Pharmacol Exp Ther (1976) 197: 517-532; Kosterlitz, et al. Endogenous opioid peptides: multiple agonists and receptors, Nature (1977) 267: 495-499) . Endogenous opioids, [Met ⁵ ]-enkephalin, [Leu ⁵ ]-enkephalin, long-acting forms of [Met ⁵ ]-enkephalin, including metorfamide and BAM-18, β-endorphin, and processed forms of dynorphin (eg dynorphin-(1-9) and shorter dynorphin peptides) also have μ receptor affinity, consistent with the possible role of each of these peptides as a natural ligand for this type of receptor, although these endogenous peptides are not selective for μ receptors. Two putative natural ligands, endomorphin 1 and 2, have also been reported that appear to act exclusively through the μ-opioid receptor and are present in the brain, although no gene, precursor protein, or any other has been identified. mechanism of their endogenous synthesis.

[00126] µ-receptors are distributed in the axial part of the central nervous system. The highest concentrations of µ-receptors are found in the thalamus, caudate nucleus and putamen, neocortex, nucleus accumbens, amygdala, interpeduncular complex, and inferior and superior colliculi (Watson et al. Autoradiographic differentiation of mu, delta and kappa receptors in the rat forebrain and midbrain, J Neurosci (1987) 7:2445-2464). The µ receptor as well as δ and κ receptors are also present in the superficial layers of the dorsal horn of the spinal cord. Small concentrations of µ-receptors are found in the periaqueductal gray matter and raphe nuclei. The role of these areas of the brain in the regulation of pain sensitivity and the implementation of the action of analgesia has been firmly established. Other physiological functions regulated by µ receptors include respiratory and cardiovascular functions, intestinal transit, nutrition, mood, thermoregulation, hormone secretion, and immune functions.

[00127] The δ opioid receptor (delta, OP ₁ or DOP) was identified using a preparation derived from the vas deferens of mice, enkephalins are generally considered to be the preferred endogenous ligands for this receptor. δ-receptors are discretely distributed throughout the central nervous system (CNS), with a significant gradient in receptor concentration from high levels in forebrain structures to relatively low levels in most regions of the hindbrain. The highest concentrations are found in the olfactory bulb, neocortex, caudate nucleus and putamen, nucleus accumbens and amygdala (Watson et al. Autoradiographic differentiation of mu, delta and kappa receptors in the rat forebrain and midbrain, J. Neurosci. (1987), 7: 2445-2464). Small concentrations of δ-receptors are found in the thalamus and hypothalamus; in more caudal areas, the interpeduncular nucleus and pons nuclei show high levels of binding in rats, but significantly lower levels in mice (Kitchen et al. Quantitative autoradiographic mapping of mu, delta and kappa-opioid receptors in kn°C kout mice lacking the mu -opioid receptor gene, Brain Res. (1997), 778: 73-88). In the spinal cord, δ-receptors are located in the dorsal horn, where they are involved in the implementation of the analgesic action of δ-agonists.

[00128] The existence of the κ opioid receptor (kappa, OP ₂ or KOP) was first indicated by the results of in vivo studies in dogs using ketocyclazocin and related drugs (Martin et al. The effects of morphine- and nalorphine-like drugs in the independent and morphine-dependent chronic spinal dog, J Pharmacol Exp Ther (1976) 197:517-532). Further studies confirmed the presence of this type of receptor in other species, including the guinea pig, the species favored in many of the early kappa opioid receptor studies. Dynorphins A and B and α-neoendorphin appear to be endogenous ligands for κ-opioid receptors, although shorter peptides derived from prodynorphin have comparable affinities for μ- and κ-receptors. In the rat and mouse, κ receptors are localized predominantly in the cerebral cortex, nucleus accumbens, brain wall, and hypothalamus (Kitchen et al. Quantitative autoradiographic mapping of mu, delta and kappa-opioid receptors in kn°C kout mice lacking the mu-opioid receptor gene, Brain Res. (1997), 778: 73-88; Watson et al. Autoradiographic differentiation of mu, delta and kappa receptors in the rat forebrain and midbrain, J. Neurosci. (1987), 7: 2445-2464) and involved in the regulation of nociception, diuresis, nutrition, neuroendocrine and immune system functions (Dhawan et al. International Union of Pharmacology. XII. Classification for opioid receptors, Pharmacol. Rev. (1996), 48: 567-592).

[00129] ORL1 receptors (also called nociceptin or orphanin FQ receptors) were identified later than all other members of the opioid receptor family. Agonist-induced internalization of the ORL1 receptor is rapid and the rate of this process is concentration dependent. Agonist stimulation also reduces the ability of the ORL1 receptor to bind to G proteins, which is required to inhibit forskolin-induced cAMP synthesis, suggesting that ORL1 undergoes similar desensitization mechanisms when compared to the other three opioid receptor subtypes.

[00130] The structure of the ORL1 receptor indicates that it is part of the opioid receptor family. Comparison of the sequence of the ORL1 receptor with those of the μ, κ, and δ receptors and other similar G protein-coupled receptors (e.g., the SOM receptor family) indicates that the ORL1 receptor is more closely related to opioid receptors than to other types of G-coupled receptors. G proteins (Birgul, et al. Reverse Physiology in drosophila: identification of a novel allatostatin-like neuropeptide and its cognate receptor structurally related to the mammalian somatostatin/galanin/opioid receptor family. EMBO J. (1999), 18: 5892- 5900). In addition, ORL1 receptor agonists induce activation of the same transduction pathways that are activated by the μ, κ, and δ receptors, and the sequence of the endogenous ORL1 receptor ligand has significant homology with the dynorphin A sequence and somewhat less with the enkephalin sequences. Accordingly, from an evolutionary point of view, the ORL1 receptor and its endogenous ligand are closely related to the μ, κ, and δ receptors.

[00131] Despite the evidence of evolutionary and functional homology, the ORL1 receptor is not an opioid receptor in terms of pharmacological prospects. The effects of activation of this receptor are not explicitly similar to those of opiates in relation to the perception of pain. The ORL1 receptor has little affinity for naloxone and most other μ, κ, or δ receptor antagonists. However, the ORL1 receptor is expressed in many functional systems in which endogenous opioids play a regulatory role. Although the functions of ORL1 are not yet fully understood, it seems very likely that the regulatory functions of ORL1 are similar to, but not identical to, those of endogenous opioid peptides. Despite these functional differences, the subcommittee concluded that the structural relationship between the ORL1 receptor and the μ, δ, and κ receptors is compelling.

[00132] Understanding the regulation of the ORL1 receptor, despite increasing research in this area, is still in its early stages compared to the results of the study of the other three opioid receptor subtypes. To date, several studies have been performed using site-directed mutagenesis, and receptor regulation in primary neurons, dorsal root ganglion, or dorsal horn neurons remains unknown.

[00133] An integral part of the effort to characterize the opioid receptor system has been the discovery of effective pure opioid receptor antagonists. Nalmefene (1A) and naltrexone (1B), competitive µ, δ and κ opioid receptor antagonists, have been used as pharmacological tools to identify and characterize the opioid system.

[00134] Nalmefene is an opioid receptor antagonist that has been available for several years as an injectable ^Revex® used to reverse opioid effects and opioid overdose. Nalmefene, as described in the literature, is also used to treat addictive substance abuse disorders such as alcohol dependence and alcohol abuse, and impulse control disorders such as pathological gambling and shopping addiction. Nalmefene is registered in Europe as Selincro and is available as an oral tablet and is taken as needed for the treatment of alcohol dependence. The name of nalmefene according to the IUPAC nomenclature is 17-cyclopropylmethyl-4,5α-epoxy-6-methylenemorphinan-3,14-diol, its structure is represented by Formula (1A).

(1A)

(1A)

[00135] Naltrexone is an opioid receptor antagonist used primarily in the treatment of alcohol dependence and opioid dependence. Naltrexone is registered in generic form as the hydrochloride salt, naltrexone hydrochloride, under the trademarks Revia ^® and Depade ^® in the form of 50 mg film-coated tablets. The once-monthly extended-release formulation of naltrexone, registered in the US as Vivitrol, has gained widespread approval for the treatment of opioid use disorders due to increased patient adherence to treatment. The IUPAC nomenclature name for naltrexone is 17-(cyclopropylmethyl)-4,5α-epoxy-3,14-dihydroxymorphinan-6-one, and its structure is represented by Formula (1B).

(1B)

(1b)

[00136] Studies have also been conducted on the use of low doses of naltrexone in patients with multiple sclerosis, autism, active Crohn's disease, AIDS (acquired immunodeficiency syndrome), rheumatoid arthritis, celiac disease, some forms of cancer and autoimmune diseases. Opioids act as cytokines, the main communication signalers of the immune system, creating immunomodulatory effects through opioid receptors on immune cells. Very low doses of naltrexone have been shown to stimulate the immune system and help fight diseases characterized by impaired immune function.

[00137] In terms of pharmacology, naltrexone blocks opioid effects by highly competitive binding to μ-opioid receptors. As a competitive antagonist, it can attenuate the suppression of euphoria induced by an opiate agonist. At the same time, clinical studies have shown that an oral dose of naltrexone of approximately 50 can block the pharmacological effects of intravenously administered doses of heroin up to 25 mg within twenty-four hours.

[00138] The mechanism of action of naltrexone in the treatment of alcoholism remains unclear, although the results of preclinical studies suggest that the endogenous opioid system is involved in this mechanism. Opioid antagonists have been shown to reduce alcohol intake in animals, and naltrexone has been shown to be effective in maintaining withdrawal symptoms in human clinical studies.

Prodrugs Opioid receptor antagonists

[00139] The use of nalmefene and naltrexone for the treatment of alcohol dependence and opioid dependence brings great benefits to society, however, there is a problem that these drugs have a very short period of action. For example, naltrexone, which is well absorbed when taken orally (approximately 96% of an oral dose is absorbed from the gastrointestinal tract), undergoes significant first-pass metabolism, and its oral bioavailability is estimated to be in the range of 5% to 40%. Naltrexone itself and its metabolite 6-β-naltrexol are believed to contribute to the activity of naltrexone. Two other minor metabolites are known: 2-hydroxy-3-methoxy-6-β-naltrexol and 2-hydroxy-3-methyl-naltrexone. The maximum concentration of naltrexone and 6-β-naltrexol in plasma is reached within one hour after oral administration; the mean half-life of naltrexone and 6-β-naltrexol is four and thirteen hours, respectively. Clinicians report that patients stop treatment too soon, even with long-acting injectable naltrexone. Therefore, ultra-long-acting opioid antagonists are needed for the treatment of substance abuse disorder.

[00140] One way to address the short duration of action of nalmefene and naltrexone is to use prodrugs that provide a sustained, sustained and controlled release of the opioid receptor antagonists nalmefene and naltrexone when administered to the body.

[00141] As used herein, the term "prodrug" is intended to mean a compound that is converted under physiological conditions to nalmefene or naltrexone. The prodrug, according to some embodiments of the present invention, is inactive when administered to a subject, but is converted in vivo to the active compound, eg, by hydrolysis. Accordingly, the term "prodrug" refers to a precursor compound that is pharmaceutically acceptable and, according to some embodiments of the present invention, lacks the pharmacological properties of nalmefene or naltrexone. The use of a prodrug often offers advantages in solubility, tissue compatibility, or delayed release in the mammalian body (see, for example, Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam).

[00142] A discussion of prodrugs can be found in Higuchi, T., et al., "Pro-drugs as Novel Delivery Systems," A.C.S. Symposium Series, Vol. 14, and in Bioreversible Carriers in Drug Design, ed. Edward B. R°C he, American Pharmaceutical Association and Pergamon Press, 1987.

[00143] The term "prodrug" is also intended to include any covalently linked carriers that release the active compound in vivo when such a prodrug is administered to a mammal. Nalmefene or naltrexone prodrugs, such as described herein, are prepared by modifying functional groups present on the active compound such that these modifications are removed to form the parent active compound. Prodrugs include compounds in which the hydroxyl group is bonded to any group which, when the prodrug of the active compound is administered to a mammal, is cleaved off to form a free hydroxyl group.

[00144] The present invention provides nalmefene and naltrexone opioid receptor antagonist prodrugs.

[00145] According to another aspect of the present invention, there is also provided a compound, or a pharmaceutically acceptable salt thereof, having the structure represented by Formula (II),

(II),

(II)

Where

X is O or CH ₂ ;

,R represents

,

where R ¹ is C ₄ -C ₁₀ alkyl or C ₄ -C ₁₀ alkenyl and n is 7-15, provided that if X is O, then n is not 7.

[00146] In some embodiments, X is O. In some embodiments, X is CH ₂ .

[00147] In some embodiments, R ¹ is C ₄ -C ₁₀ alkyl or C ₄ -C ₁₀ alkenyl.

[00148] In some embodiments, R ¹ is C ₄ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₅ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₆ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₆ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₉ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₄ alkyl. According to some embodiments of the invention, R ¹ is C ₅ alkyl. According to some embodiments of the invention, R ¹ is C ₆ alkyl. According to some embodiments of the invention, R ¹ is C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₁₀ alkyl.

[00149] In some embodiments, R ¹ is C ₄ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₅ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₆ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₇ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₆ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₇ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₇ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₉ -C ₁₀ alkenyl. In some embodiments, R ¹ is C ₄ alkenyl. In some embodiments, R ¹ is C ₅ alkenyl. In some embodiments, R ¹ is C ₆ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ alkenyl. In some embodiments, R ¹ is C ₈ alkenyl. In some embodiments, R ¹ is C ₉ alkenyl. In some embodiments, R ¹ is C ₁₀ alkenyl.

[00150] According to some embodiments of the invention, n has a value of 7-15. According to some embodiments of the invention, n has a value of 7-8. According to some embodiments of the invention, n has a value of 7-9. According to some embodiments of the invention, n has a value of 7-10. According to some embodiments of the invention, n has a value of 7-11. According to some embodiments of the invention, n has a value of 7-12. According to some embodiments of the invention, n has a value of 7-13. According to some embodiments of the invention, n has a value of 7-14. According to some embodiments of the invention, n has a value of 8-9. According to some embodiments of the invention, n has a value of 8-10. According to some embodiments of the invention, n has a value of 8-11. According to some embodiments of the invention, n has a value of 8-12. According to some embodiments of the invention, n has a value of 8-13. According to some embodiments of the invention, n has a value of 8-14. According to some embodiments of the invention, n has a value of 8-15. According to some embodiments of the invention, n has a value of 9-10. According to some embodiments of the invention, n has a value of 9-11. According to some embodiments of the invention, n has a value of 9-12. According to some embodiments of the invention, n has a value of 9-13. According to some embodiments of the invention, n has a value of 9-14. According to some embodiments of the invention, n has a value of 9-15. According to some embodiments of the invention, n has a value of 10-11. According to some embodiments of the invention, n has a value of 10-12. According to some embodiments of the invention, n has a value of 10-13. According to some embodiments of the invention, n has a value of 10-14. According to some embodiments of the invention, n has a value of 10-15. According to some embodiments of the invention, n has a value of 11-12. According to some embodiments of the invention, n has a value of 11-13. According to some embodiments of the invention, n has a value of 11-14. According to some embodiments of the invention, n has a value of 11-15. According to some embodiments of the invention, n has a value of 12-13. According to some embodiments of the invention, n has a value of 12-14. According to some embodiments of the invention, n has a value of 12-15. According to some embodiments of the invention, n has a value of 13-14. According to some embodiments of the invention, n has a value of 13-15. According to some embodiments of the invention, n has a value of 14-15. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, n is 11. In some embodiments, n is 12. In some embodiments, n is 13. In some embodiments, n is 14. In some embodiments, n is 15.

[00151] According to another aspect of the present invention, there is also provided a compound, or a pharmaceutically acceptable salt thereof, having the structure represented by Formula (IIa),

(IIa),

(IIa),

Where

X is O or CH ₂ ;

R represents

,

,

where R ¹ is C ₄ -C ₁₀ alkyl or C ₄ -C ₁₀ alkenyl and n is 9-15.

[00152] In some embodiments, X is O. In some embodiments, X is CH ₂ .

[00153] In some embodiments, R ¹ is C ₄ -C ₁₀ alkyl or C ₄ -C ₁₀ alkenyl.

[00154] In some embodiments, R ¹ is C ₄ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₅ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₆ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₆ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₉ -C ₁₀ alkyl. According to some embodiments of the invention, R ¹ is C ₄ alkyl. According to some embodiments of the invention, R ¹ is C ₅ alkyl. According to some embodiments of the invention, R ¹ is C ₆ alkyl. According to some embodiments of the invention, R ¹ is C ₇ alkyl. According to some embodiments of the invention, R ¹ is C ₈ alkyl. According to some embodiments of the invention, R ¹ is C ₉ alkyl. According to some embodiments of the invention, R ¹ is C ₁₀ alkyl.

[00155] In some embodiments, R ¹ is C ₄ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₅ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₆ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₇ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₄ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₆ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₇ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₅ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₇ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₆ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₈ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₉ alkenyl. According to some embodiments of the invention, R ¹ is C ₈ -C ₁₀ alkenyl. According to some embodiments of the invention, R ¹ is C ₉ -C ₁₀ alkenyl. In some embodiments, R ¹ is C ₄ alkenyl. In some embodiments, R ¹ is C ₅ alkenyl. In some embodiments, R ¹ is C ₆ alkenyl. According to some embodiments of the invention, R ¹ is C ₇ alkenyl. In some embodiments, R ¹ is C ₈ alkenyl. In some embodiments, R ¹ is C ₉ alkenyl. In some embodiments, R ¹ is C ₁₀ alkenyl.

[00156] According to some embodiments of the invention, n has a value of 9-15. According to some embodiments of the invention, n has a value of 9-10. According to some embodiments of the invention, n has a value of 9-11. According to some embodiments of the invention, n has a value of 9-12. According to some embodiments of the invention, n has a value of 9-13. According to some embodiments of the invention, n has a value of 9-14. According to some embodiments of the invention, n has a value of 10-11. According to some embodiments of the invention, n has a value of 10-12. According to some embodiments of the invention, n has a value of 10-13. According to some embodiments of the invention, n has a value of 10-14. According to some embodiments of the invention, n has a value of 10-15. According to some embodiments of the invention, n has a value of 11-12. According to some embodiments of the invention, n has a value of 11-13. According to some embodiments of the invention, n has a value of 11-14. According to some embodiments of the invention, n has a value of 11-15. According to some embodiments of the invention, n has a value of 12-13. According to some embodiments of the invention, n has a value of 12-14. According to some embodiments of the invention, n has a value of 12-15. According to some embodiments of the invention, n has a value of 13-14. According to some embodiments of the invention, n has a value of 13-15. According to some embodiments of the invention, n has a value of 14-15. In some embodiments, n is 9. In some embodiments, n is 10. In some embodiments, n is 11. In some embodiments, n is 12. In some embodiments, n is 13. In some embodiments, n is 14. In some embodiments, n is 15.

[00157] According to another aspect of the present invention, there is also provided a compound, or a pharmaceutically acceptable salt thereof, having the structure represented by Formula (III),

(III),

(III)

Where

X is O or CH ₂ ;

R is selected from

-[CH(R ³ )O]zR ⁴ ;

-[CH(R ³ )O]zC(=O)OR ⁴ ;

-[CH(R ³ )O]zC(=O)NR ⁴ R ⁵ and

-[CH(R ³ )O]zP(=O)(OR ⁴ )(OR ⁵ ),

where z is 1, 2, 3, 4, 5, 6, or 7;

R ³ represents a hydrogen atom, a halogen atom, alkyl, alkenyl, cycloalkylalkyl or aryl;

each R ⁴ and R ⁵ is independently selected from hydrogen, alkyl, alkenyl, cycloalkylalkyl or aryl.

[00158] In some embodiments, X is O. In some embodiments, X is CH ₂ .

[00159] In some embodiments, z is 1. In some embodiments, z is 2. In some embodiments, z is 3. In some embodiments, z is 4. In some embodiments, z is value 5. In some embodiments, z is 6. In some embodiments, z is 7. In some embodiments, z is 1 or 2. In some embodiments, z is 2 or 3. In some embodiments, z is 2 or 3. implementation of the invention z has a value of 1, 2 or 3.

[00160] According to some embodiments of the invention, R ³ represents a hydrogen atom, a halogen atom, or alkyl. According to some embodiments of the invention, R ³ is alkyl. According to some embodiments of the invention R ³ represents a hydrogen atom. According to some embodiments of the invention R ³ represents a hydrogen atom, a halogen atom, alkyl, cycloalkylalkyl or aryl. According to some embodiments of the invention R ³ represents a hydrogen atom, a halogen atom, cycloalkylalkyl or aryl. According to some embodiments of the invention R ³ represents a halogen atom. In some embodiments, the halogen atom is a fluorine atom.

[00161] According to some embodiments of the invention, each R ⁴ and R ⁵ is independently selected from alkyl or aryl. According to some embodiments of the invention, each R ⁴ and R ⁵ is independently selected from alkyl. According to some embodiments of the invention, each R ⁴ and R ⁵ is independently selected from hydrogen or alkyl. According to some embodiments of the invention, alkyl is C ₁₀ -C ₁₈ alkyl. In some embodiments, alkyl is C ₅ -C ₉ alkyl. In some embodiments, alkyl is C ₁ -C ₄ alkyl. In some embodiments, alkyl is C ₉ -C ₁₃ alkyl. In some embodiments, alkyl is C ₁₀ -C ₁₂ alkyl. In some embodiments, alkyl is C ₁₀ alkyl. In some embodiments, alkyl is C ₁₁ alkyl. In some embodiments, alkyl is C ₁₂ alkyl.

[00162] In some embodiments, R is -[CH(R ³ )O]zR ⁴ . According to some embodiments of the invention, R is -[CH(R ³ )O]zC(=O)OR ⁴ . In some embodiments, R is -[CH(R ³ )O]zC(=O)NR ⁴ R ⁵ . In some embodiments, R is -[CH(R ³ )O]zP(=O)(OR ⁴ )(OR ⁵ ). According to some variants of the invention, R represents -[CH(R ³ )O]zC(=O)OR ⁴ where R ³ represents a hydrogen atom and R ⁴ represents C ₉ -C ₁₃ alkyl. In some embodiments, R is -[CH(R ³ )O]zC(=O)OR ⁴ , where R ³ is hydrogen and R ⁴ is C ₁₀ -C ₁₂ alkyl. In some embodiments, R is -[CH(R ³ )O]zC(=O)OR ⁴ , where R ³ is hydrogen and R ⁴ is C ₁₀ alkyl. According to some variants of the invention, R represents -[CH(R ³ )O]zC(=O)OR ⁴ where R ³ represents a hydrogen atom and R ⁴ represents C ₁₁ alkyl. According to some variants of the invention, R represents -[CH(R ³ )O]zC(=O)OR ⁴ where R ³ represents a hydrogen atom and R ⁴ represents C ₁₂ alkyl.

[00163] According to some embodiments of the present invention, the opioid receptor antagonist prodrug compound described herein has the structure shown in Table 1.

TABLE 1

Example
chemical
synthesis R X chemical name 1

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl ((E)-octadec-9-en-1-yl) carbonate 2

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl undecyl carbonate 3

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl undecyl carbonate 4

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl undecyl carbonate 5

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl undecyl carbonate 6

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl dodecyl carbonate 7

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl ((E)-octadec-9-en-1-yl) carbonate 8

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (E)-octadec-9-enoate 9

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl (E)-octadec-9-enoate 10

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl decyl carbonate eleven

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl dodecyl carbonate 12

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl stearate 13

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (Z)-docose-13-enoate 14

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl docosanoate 15

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (E)-octadec-9-enoate 16

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl eicosanoate 17

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl octyl carbonate 18

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl decyl carbonate 19

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl hexadecyl carbonate 20

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (9Z,12Z,15Z)-octadeca-9,12,15-trienoate 21

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl hexadecyl carbonate 22

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (Z)-docose-13-enoate 23

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl octyl carbonate 24

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl dodecyl carbonate 25

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl tetradecyl carbonate 26

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl (E)-octadec-9-enoate 27

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl tetradecyl carbonate 28

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl eicosyl carbonate 29

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl dodecyl carbonate thirty

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl tridecyl carbonate 31

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl tetradecyl carbonate 32

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl pentadecyl carbonate 33

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl octadecyl carbonate 34

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl hexadecyl carbonate 35

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl decyl carbonate 36

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl oleate 37

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (9Z,12Z)-octadeca-9,12-dienoate 38

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl 3,3-dimethylbutanoate 39

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl 3-cyclopentylpropanoate 40

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl tert-butyl carbamate 41

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl oleate 42

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl 3,3-dimethylbutanoate 43

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl 3-cyclopentylpropanoate 44

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl dodecanoate 45

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl tetradecanoate 46

CH ₂ (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl hexadecanoate 47

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl hexadecyl carbonate 48

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl dodecanoate 49

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl hexadecanoate

[00164] According to some embodiments of the present invention, the opioid receptor antagonist prodrug compound described herein has the structure shown in Table 2.

TABLE 2

Example R X chemical name 50

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl eicosanoate 51

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl docosanoate 52

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (9Z,12Z)-octadeca-9,12,15-trienoate 53

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (9Z,12Z)-octadeca-9,12-di.enoate 54

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl stearate 55

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl palmitate 56

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl palmitate 57

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl tetradecanoate 58

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl pentadecanoate 59

CH ₂ (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl dodecanoate 60

O (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl)oxy)methyl tetradecanoate 61 H CH ₂ Nalmefene 62 H O Naltrexone

Getting Connections

[00165] the Compounds used in the reactions described in this description, obtained in accordance with the methods of organic synthesis, known to experts in the art, from commercially available chemicals and/or from compounds described in the chemical literature. "Commercially available chemicals" are obtained from standard commercial sources, including Acros Organics (Pittsburgh, PA), Aldrich Chemical (Milwaukee, WI; including Sigma Chemical and Fluka), Apin Chemicals Ltd. (Milton Park, United Kingdom), Av°C ado Research (Lancashire, United Kingdom), BDH Inc. (Toronto, Canada), Bionet (Cornwall, United Kingdom), Chemservice Inc. (Westchester, PA), Crescent Chemical Co. (Hopag, NY), Eastman Organic Chemicals, Eastman Kodak Company (Rochester, NY), Fisher Scientific Co. (Pittsburgh, PA), Fisons Chemicals (Leicestershire, UK), Frontier Scientific (Logan, UT), ICN Biomedicals, Inc. (Costa Mesa, CA), Key Organics (Cornwall, UK), Lancaster Synthesis (Wyndham, NH), Maybridge Chemical Co. Ltd. (Cornwall, United Kingdom), Parish Chemical Co. (Orem, Utah), Pfaltz & Bauer, Inc. (Waterbury, CT), Polyorganix (Houston, TX), Pierce Chemical Co. (Rockford, IL), Riedel de Haen AG (Hannover, Germany), Spectrum Quality Product, Inc. (New Brunswick, NJ), TCI America (Portland, Oregon), Trans World Chemicals, Inc. (Rockville, Maryland) and Wako Chemicals USA, Inc. (Richmond, Virginia).

[00166] Suitable reference books and monographs, which provide detailed methods for the synthesis of reagents that can be used to obtain the compounds described in this application, or provide links to articles that describe the preparation of these reagents, include, for example, "Synthetic Organic Chemistry" , John Wiley & Sons, Inc., New York; S.R. Sandler et al., "Organic Functional Group Preparations," ^2nd Ed., Academic Press, New York, 1983; HO House, "Modern Synthetic Reactions", ^2nd Ed., WA Benjamin, Inc. Menlo Park, Calif. 1972; TL Gilchrist, "Heter°C yclic Chemistry", ^2nd Ed., John Wiley & Sons, New York, 1992; J. March, "Advanced Organic Chemistry: Reactions, Mechanisms and Structure", ^4th Ed., Wiley-Interscience, New York, 1992 , described in this application, or references are made to articles that describe the preparation of these reagents, include, for example, Fuhrhop, J. and Penzlin G. "Organic Synthesis: Concepts, Methods, Starting Materials", Second, Revised and Enlarged Edition (1994 ) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R.V. "Organic Chemistry, An Intermediate Text" (1996) Oxford University Press, ISBN 0-19-509618-5; Lar°C k, RC "Comprehensive Organic Transformations: A Guide to Functional Group Preparations" ^2nd Edition (1999) Wiley-VCH, ISBN: 0-471-19031-4; March, J. "Advanced Organic Chemistry: Reactions, Mechanisms, and Structure" ^4th Edition (1992) John Wiley & Sons, ISBN: 0-471-60180-2; Otera, J. (ed.) "Modern Carbonyl Chemistry" (2000) Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. "Patai's 1992 Guide to the Chemistry of Functional Groups" (1992) Interscience ISBN: 0-471-93022-9; Solomons, TWG "Organic Chemistry" ^7th Edition (2000) John Wiley & Sons, ISBN: 0-471-19095-0; Stowell, JC, "Intermediate Organic Chemistry" 2nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2; "Industrial Organic Chemicals: Starting Materials and Intermediates: An Ullmann's Encyclopedia" (1999) John Wiley & Sons, ISBN: 3-527-29645-X, in 8 volumes; "Organic Reactions" (1942-2000) by John Wiley & Sons, in more than 55 volumes, and "Chemistry of Functional Groups" by John Wiley & Sons, in 73 volumes.

[00167] Specifically used and similar reagents, if necessary, can be found through the indexes of known chemical reagents issued by the Chemical Abstract Service of the American Society of Chemists (American Chemical S ° C iety), which are available in most public and university libraries, and through online databases (contact the American Society of Chemists, Washington DC for more details). Chemicals that are known but not listed for sale can be synthesized by custom chemical synthesis firms if desired, with many of the standard chemical vendors (such as those listed above) providing custom synthesis services. As reference for the preparation and selection of pharmaceutical salts of the compounds described herein for use as opioid receptor antagonist prodrugs, P. H. Stahl & C. G. Wermuth "Handbook of Pharmaceutical Salts", Verlag Helvetica Chimica Acta, Zurich, 2002.

Pharmaceutical Compositions

[00168] According to some embodiments of the present invention, the opioid receptor antagonist prodrug compound described herein is administered as a pure chemical. According to other embodiments of the present invention, the opioid receptor antagonist prodrug compound described herein is combined with a pharmaceutically acceptable or acceptable carrier (also referred to herein as a pharmaceutically acceptable (or acceptable) excipient, a physiologically acceptable (or acceptable) excipient, or a physiologically acceptable (or acceptable) carrier) selected based on the route of administration used and standard pharmaceutical practice, for example, as described in Remington: The Science and Practice of Pharmacy (Gennaro, 21 ^st Ed. Mack Pub. Co., Easton , PA (2005)).

[00169] The present invention provides a pharmaceutical composition comprising at least one opioid receptor antagonist prodrug compound, or stereoisomer, pharmaceutically acceptable salt, hydrate, solvate, or N-oxide thereof, together with one or more pharmaceutically acceptable carriers. The carrier(s) (or excipient(s)) is acceptable or appropriate if the carrier is compatible with the other ingredients of the composition and safe for the recipient (ie subject) of the composition.

[00170] According to one embodiment of the present invention, there is provided a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound represented by any of Formulas (II), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt of these connections.

[00171] According to some embodiments of the present invention, the opioid receptor antagonist prodrug compound represented by any of Formulas (II), (IIa) or (III), or the compound shown in Table 1, is substantially pure, i.e. contains less about 5%, or less than about 1%, or less than about 0.1%, other organic small molecules, such as unreacted intermediates or synthetic by-products, which are formed, for example, in one or more steps of the synthetic procedure.

[00172] Suitable oral dosage forms include, for example, tablets, pills, sachets or capsules of hard or soft gelatin, methylcellulose, or other suitable substance readily soluble in the gastrointestinal tract. According to some embodiments of the present invention, suitable non-toxic solid carriers are used, which include, for example, mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate, and the like pharmaceutical grade substances. (See, for example, Remington: The Science and Practice of Pharmacy (Gennaro, 21 ^st Ed. Mack Pub. Co., Easton, PA (2005)).

[00173] According to some embodiments of the present invention, the opioid receptor antagonist prodrug compound represented by any of Formulas (II), (IIa) or (III), or the compound shown in Table 1, is formulated for administration by injection. In some cases, this injectable preparation is an aqueous preparation. In some cases, the injectable formulation is a non-aqueous formulation. In some cases, this injectable preparation is an oil-based preparation such as sesame oil, cottonseed oil or the like.

[00174] The dose of a composition containing at least one opioid receptor antagonist prodrug compound, such as described herein, depends on the condition of the patient (e.g., human), i.e., general health, age, and other factors.

[00175] Pharmaceutical compositions are administered by a route appropriate for the disease in need of treatment (or prevention). The appropriate dosage and the appropriate duration and frequency of administration will generally be determined by such factors as the condition of the patient, the type and severity of the disease the patient is suffering from, the particular form of the active ingredient, and the route of administration. In general, a suitable dosage and treatment regimen will provide sufficient amount of the composition(s) to produce a therapeutic and/or prophylactic effect (e.g., a better clinical outcome, such as a higher rate of complete or partial remission, or a longer remission stage and/or a higher overall survival, or reduction in symptoms.Optimal doses are usually determined using experimental models and/or clinical studies.The optimal dose depends on the body weight, weight, or blood volume of the patient.

Doses and treatment regimens

[00176] According to some embodiments of the present invention, the pharmaceutical compositions described in this application are administered for therapeutic purposes. In some embodiments, the pharmaceutical composition is administered once a day, twice a day, three times a day, four times a day, or more frequently. The pharmaceutical composition is administered daily, once a day, every other day, two days a week, three days a week, four days a week, five days a week, once a week, once every two weeks, two weeks a month, three weeks a month , once a month, twice a month, three times a month, or with another greater or lesser frequency of administration; the pharmaceutical composition can also be administered once every 2 months, once every 3 months, once every 4 months, once every 5 months, once every 6 months, once a year, or less or more than the above-mentioned administration intervals. The pharmaceutical composition is administered for at least 1 week, 2 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 18 months, 2 years, 3 years or more.

[00177] In the event that the patient's condition does not improve, at the discretion of the physician, the administration of the composition does not stop; alternatively, the dose of the administered composition is temporarily reduced or the administration is temporarily suspended for some time (ie, a "drug holiday" is administered). In some cases, the duration of the drug holiday is from 2 days to 1 year, including, by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days , 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, 365 days or 366 days. Dose reduction during drug holidays is 10%-100% including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100%.

[00178] After the onset of improvement in the patient's condition, if necessary, a maintenance dose is administered. The dose or frequency of administration, or both, can then be adjusted, depending on the symptoms, to a level at which improvement is maintained over the course of the disease, disorder, or condition.

[00179] According to some embodiments of the present invention, the amount of an opioid antagonist prodrug compound administered varies depending on factors such as the particular compound, disease severity, characteristics (e.g., weight) of the subject or recipient in need of treatment, but nonetheless usually determined, as is customary in the art, in accordance with the specific circumstances surrounding a particular case, including, for example, the particular agent administered, the route of administration, and the characteristics of the subject or recipient in need of treatment. In some instances, the desired dose is conveniently presented as a single dose or as separate doses administered simultaneously (or over a short period of time) or at suitable intervals, such as two, three, four or more sub-doses administered over a period of time. days.

[00180] In some embodiments, the amount of opioid antagonist prodrug compound administered is typically in the range of about 0.02 mg to about 5000 mg per dose. (Note: for all prodrugs, weight is expressed in base grouping equivalents.) In some embodiments of the present invention, the amount of opioid antagonist prodrug compound administered is in the range of about 1 mg to about 5000 mg per dose. In some embodiments, the amount of opioid antagonist prodrug compound administered is in the range of about 10 mg to about 1600 mg per dose. The desired dose may conveniently be presented as a single dose or as separate doses administered simultaneously (or over a short period of time) or at suitable intervals, such as two, three, four or more sub-doses administered throughout the day. .

[00181] According to some embodiments of the present invention, suitable daily doses of the opioid receptor antagonist prodrug compound described herein are in the range of about 0.01 mg/kg to about 30 mg/kg. According to one of the embodiments of the present invention, daily doses are in the range from about 0.1 mg/kg to about 165 mg/kg. The daily dose indicated for administration to large mammals, including but not limited to humans, ranges from about 0.5 mg to about 1000 mg, and for convenience, is administered as a single dose or as divided doses. Unit dosage forms suitable for intramuscular administration include from about 1 to about 5000 mg of the active ingredient. According to one embodiment of the present invention, the dosage unit is about 10 mg, about 50 mg, about 100 mg, about 200 mg, about 500 mg, about 1000 mg, about 2000 mg, about 2500 mg, about 4000 mg, or about 5000 mg. .

[00182] The above ranges are given for illustrative purposes only, as there are a huge number of variables in an individual treatment regimen, and it is not uncommon for significant deviations from these recommended values. Such dosages may vary depending on a number of variables including, but not limited to, the activity of the compound being used, the disease or condition being treated, the route of administration, the individual needs of the subject, the severity of the disease or condition being treated, and the judgment of the attending physician.

conduct disorder treatment

[00183] In some embodiments, the invention describes a method of treating one or more medical conditions in a subject in need of such treatment, comprising administering an opioid receptor antagonist compound described herein to the subject in need.

[00184] According to some embodiments of the present invention, the medical condition is selected from the group consisting of opioid dependence, alcohol dependence, drug addiction, polydrug addiction, and pain.

[00185] In some embodiments, the present invention describes an opioid receptor antagonist compound for use in reducing opioid consumption in an opioid dependent patient.

[00186] In some embodiments, the present invention describes an opioid receptor antagonist compound for use in reducing alcohol consumption in a patient with alcohol dependence, pathological gambling and oniomania, or other compulsive behavior disorders.

[00187] The present invention provides a method for treating opioid dependence in a patient in need of such treatment, comprising administering a pharmaceutical composition containing a compound of Formula (II), (IIa) or (III), or a compound shown in Table 1, or a pharmaceutically acceptable salt said compounds, and a pharmaceutically acceptable excipient. The present invention provides a method, characterized in that the pharmaceutical composition is administered orally. The present invention provides a method, characterized in that the pharmaceutical composition is administered by injection. The present invention provides a method, characterized in that the pharmaceutical composition is administered by intramuscular injection. The present invention provides a method characterized in that the intramuscular injection is a depot injection. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of 2 days to 3 months. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 2 days. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 4 days. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 7 days. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 10 days. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 1 week. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 2 weeks. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 3 weeks. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 4 weeks. The present invention provides a method wherein the depot injection provides a therapeutically effective concentration over a period of approximately 5 weeks. The present invention provides a method wherein the depot injection provides a therapeutically effective concentration over a period of approximately 6 weeks. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 1 month. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 2 months. The present invention provides a method wherein the depot injection provides a therapeutically effective concentration over a period of approximately 3 months. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 4 months. The present invention provides a method characterized in that the depot injection provides a therapeutically effective concentration over a period of approximately 5 months. The present invention provides a method wherein the depot injection provides a therapeutically effective concentration over a period of approximately 6 months or more.

[00188] The present invention provides a method for treating opioid dependence in a patient in need of such treatment, comprising administering a pharmaceutical composition comprising a compound of Table 3, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

TABLE 3

R X chemical name

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl palmitate

O (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (9Z,12Z)-octadeca-9,12-dienoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl palmitoleate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl myristoleate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl (Z)-hexadec-6-enoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl decanoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl undecanoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl dodecanoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl tridecanoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl tetradecanoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl pentadecanoate

O (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl stearate

[00189] Other embodiments and applications of the present invention are obvious to a person skilled in the art in light of this description. The following examples are provided solely as an illustration of various embodiments of the invention and should in no way be construed as limiting the invention.

examples

I. Chemical synthesis

[00190] Unless otherwise noted, reagents and solvents were used as received from commercial suppliers. For moisture and/or oxygen sensitive chemical transformations, anhydrous solvents and oven-dried glassware were used. The yield of the reactions was not optimized. Reaction times are approximate and optimized. Column chromatography and thin layer chromatography (TLC) were performed on silica gel unless otherwise noted. The spectra are expressed in ppm (δ), and the spin-spin coupling constants, J, are expressed in hertz. For proton spectra, the solvent peak was used as a reference peak.

[00191] In some embodiments, the opioid antagonist prodrug compounds described herein are synthesized as described in the following examples.

[00192] General Scheme 1 for the synthesis of nalmefene prodrugs

Scheme 1

[00193] General Scheme 2 for the Synthesis of Naltrexone Prodrugs

Scheme 2

[00194] Example 1: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl ((E)-octadec-9-en-1-yl) carbonate

[00195] To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2 -e]isoquinoline-4a,9-diol (8 g, 21.28 mmol, 1 eq., HCl) and H ₂ O (100 ml) were added K ₂ CO ₃ (8.82 g, 63.85 mmol, 3 equiv.) at one time at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. Tetrabutylammonium sulfate (24.73 g, 21.28 mmol, 24.49 ml, 50% solution, 1 eq.) and DCM (100 ml) were mixed, and then this mixture was added to the previously obtained mixture. Iodomethyl (E)-octadec-9-en-1-yl carbonate (14.44 g, 31.92 mmol, 1.5 eq) prepared according to the procedure described in Example 42B was added and the mixture was stirred for within 12 hours. The residue was concentrated in vacuo to remove DCM, then dissolved in saturated NaHCO ₃ solution (100 ml). The aqueous phase was extracted with ethyl acetate (600 ml; 200 ml * 3). The combined organic phases were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=20/1-1/1). The residue was further purified by preparative HPLC using MeOH as solvent, standard reverse phase chromatography separation technique, and TFA separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (900 ml; 300 ml * 3). The combined organic phases were washed with brine (200 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl [(E)-octadec-9-enyl] carbonate (5 g, 7.46 mmol, yield: 35.03%) was obtained as a yellow oil. M+H ⁺ = 665.5 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 1.

[00196] Example 2: Step 2A: Synthesis of (4-nitrophenyl) undecyl carbonate

[00197] To a mixture of undecan-1-ol (40 g, 232.14 mmol, 1 eq.) in DCM (600 ml) was added TEA (46.98 g, 464.29 mmol, 64.62 ml, 2 eq. ), and (4-nitrophenyl) carbonochloridate (70.19 g, 348.22 mmol, 1.5 eq.) was added portionwise under N ₂ atmosphere. This mixture was stirred for 12 h at 25°C. The reaction mixture was then concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography. The compound (4-nitrophenyl)undecyl carbonate (33.95 g, 100.62 mmol, yield: 43.34%) was obtained as a yellow solid.

[00198] Step 2B: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl undecyl carbonate

[00199] To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3 ,2-e]isoquinolin-7-one (15 g, 39.70 mmol, 1 eq., HCl) and DCM (150 mL) TEA (12.05 g, 119.09 mmol, 16.58 mL, 3 equiv.) at one time at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. (4-nitrophenyl) undecyl carbonate (26.79 g, 79.39 mmol, 2 eq.) and DCM (150 ml) were mixed and then this mixture was added to the previously obtained mixture, the resulting mixture was stirred for 12 h at 25° C. The residue was concentrated in vacuo to remove DCM, then dissolved in saturated NaHCO ₃ solution. The aqueous phase was extracted with ethyl acetate (200 ml*3). The combined organic phases were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=40:1–1:1). The residue was further purified by preparative HPLC using MeOH as solvent, standard reverse phase chromatography separation technique, and TFA separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (200 ml * 3). The combined organic phases were washed with brine (500 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]undecyl carbonate (7.91 g, 14.63 mmol, yield: 36.85%) was obtained as a yellow oil. M+H ⁺ = 540.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 2.

[00200] Example 3: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl undecyl carbonate

[00201] To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2 -e]isoquinoline-4a,9-diol (15 g, 39.91 mmol, 1 eq, HCl) and DCM (150 mL) TEA (12.11 g, 119.72 mmol, 16.66 mL, 3 equiv.) at one time at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. (4-nitrophenyl)undecyl carbonate (26.93 g, 79.81 mmol, 2 eq.) and DCM (150 ml) were mixed and then this mixture was added to the previously obtained mixture, the resulting mixture was stirred for 12 h at 25° C. The mixture was diluted with H ₂ O (800 ml), extracted using DCM (300 ml * 3). The organic phase was washed with brine (300 ml), dried over Na ₂ SO ₄ , filtered, and the resulting filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=40/1-1/1). The residue was further purified by preparative HPLC using MeOH as solvent, standard reverse phase chromatography separation technique, and TFA separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (200 ml * 3). The combined organic phases were washed with brine (500 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]undecyl carbonate (11.40 g, 21.14 mmol, yield: 52.97%) was obtained as a yellow oil. M+H ⁺ = 538.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 3.

[00202] Example 4: Step 4A: Synthesis of chloromethyl undecyl carbonate

[00203] To a mixture of undecan-1-ol (80 g, 464.29 mmol, 1 eq.) and pyridine (73.45 g, 928.58 mmol, 74.95 ml, 2 eq.) in DCM (600 ml ) was added dropwise chloroformic acid chloromethyl ether (119.73 g, 928.58 mmol, 82.57 ml, 2 eq.) at 0°C under N ₂ atmosphere. This mixture was stirred for 12 h at 25°C. Then the reaction mixture was extracted using DCM (1500 ml; 500 ml * 3). The organic phase was separated, washed with brine 30 ml (150 ml * 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=50/1–1:1). The compound chloromethyl undecyl carbonate (80 g, 302.13 mmol, yield: 65.07%) was obtained as a yellow oil.

[00204] Step 4B: Synthesis of iodomethyl undecyl carbonate

[00205] NaHCO ₃ (11.42 g, 135.96 mmol, 5.29 ml, 1 .2 eq.) and NaI (20.38 g, 135.96 mmol, 1.2 eq.) at 25°C under N ₂ atmosphere. This mixture was stirred in the dark for 12 hours at 25°C. The reaction mixture was then partitioned between EtOAc (400 ml) and H ₂ O (400 ml). The organic phase was separated, washed with brine (80 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0). The compound iodomethyl undecyl carbonate (60 g, yield: 74.33%) was obtained as a yellow oil.

[00206] Step 4C: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl undecyl carbonate

[00207] To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3 ,2-e]isoquinolin-7-one (8 g, 21.17 mmol, 1 eq, HCl) and H ₂ O (40 ml) were added in one go K ₂ CO ₃ (8.78 g, 63.52 mmol, 3 equiv.) at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. Then tetrabutylammonium sulfate (24.60 g, 21.17 mmol, 24.36 ml, 50% solution, 1 eq.) in DCM (40 ml) at 25°C was added in one go. Iodomethyl undecyl carbonate (15.08 g, 42.34 mmol, 2 eq.) was then added to this mixture, and the resulting mixture was stirred for 11.5 hours at 25°C. Then the reaction mixture was partitioned between DCM (200 ml; 100 ml * 2) and H ₂ O (100 ml). The organic phase was separated, washed with brine (40 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=50/1–1:1). It was then further purified by preparative HPLC using MeOH as the solvent, standard reverse phase chromatography as the separation technique, and TFA as the separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (400 ml * 3), washed with brine (300 ml), dried with anhydrous Na ₂ SO ₄ , filtered and concentrated under vacuum. Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl undecyl carbonate (6.9 g) was obtained as a yellow oil. M+H ⁺ = 570.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 4.

[00208] Example 5: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl undecyl carbonate

[00209] To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2 -e]isoquinoline-4a,9-diol (8 g, 21.28 mmol, 1 eq, HCl) and H ₂ O (40 ml) were added in one go K ₂ CO ₃ (8.82 g, 63.85 mmol, 3 equiv.) at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. Then tetrabutylammonium sulfate (24.73 g, 21.28 mmol, 24.49 ml, 50% solution, 1 eq.) in DCM (40 ml) at 25°C was added in one go. Then iodomethyl undecyl carbonate (15.16 g, 42.57 mmol, 2 eq.) was added to this mixture, this mixture was stirred for 11.5 h at 25°C. Then the reaction mixture was partitioned between DCM (200 ml; 100 ml * 2) and H ₂ O (100 ml). The organic phase was separated, washed with brine (50 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=50/1–1:1). It was then further purified by preparative HPLC using MeOH as the solvent, standard reverse phase chromatography as the separation technique, and TFA as the separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (400 ml * 3), washed with brine (300 ml), dried with anhydrous Na ₂ SO ₄ , filtered and concentrated under vacuum. Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl undecyl carbonate (5.9 g) was obtained as a yellow oil. M+H ⁺ = 568.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 5.

[00210] Example 6: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl dodecyl carbonate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 6. Briefly, to a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinoline-4a,9-diol (2.5 g, 6.65 mmol, 1 eq., HCl) and DCM (10 mL) TEA (2.02 g, 19.95 mmol, 2.78 ml, 3 eq.) and dodecyl carbonochloridate (2.48 g, 9.98 mmol, 1.5 eq.). This mixture was stirred for 1 hour at -10°C and then heated to 25°C for 4 hours under N ₂ atmosphere. The reaction mixture was then concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 5/1 to 1:10). Compound [(4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl dodecyl carbonate] (purity 98.570%) was obtained as a yellow oil (1.5 g, yield: 40.56%).

[00211] Example 7: Step 7A: Synthesis of chloromethyl (E)-octadec-9-en-1-yl carbonate

[00212] To a mixture of (E)-octadec-9-en-1-ol (22 g, 81.94 mmol, 1 eq) and chloromethyl ester chloroformic acid (21.13 g, 163.89 mmol, 14.57 ml, 2 eq.) in DCM (200 ml) was added dropwise pyridine (16.20 g, 204.86 mmol, 16.54 ml, 2.5 eq.) at 0°C under N ₂ atmosphere. This reaction mixture was stirred for 12 h at 25° C. under N ₂ . Then the reaction mixture was quenched by adding H ₂ O (400 ml) and was extracted using DCM (400 ml * 1). The combined organic layers were washed with brine (300 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give an oil. This residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0) to give the product. The compound chloromethyl (E)-octadec-9-en-1-yl carbonate (50 g, 138.52 mmol, yield: 84.52%) was obtained as a colorless oil.

[00213] Step 7B: Synthesis of iodomethyl (E)-octadec-9-en-1-yl carbonate

[00214] To a mixture of chloromethyl (E)-octadec-9-en-1-yl carbonate (30 g, 83.11 mmol, 1 eq.) and NaI (18.69 g, 124.67 mmol, 1.5 eq. .) in acetone (300 ml) NaHCO ₃ (8.38 g, 99.73 mmol, 3.88 ml, 1.2 eq.) was added in one go at 25° C. under N ₂ atmosphere. This mixture was stirred for 12 h at 25°C. The reaction mixture was then concentrated under reduced pressure to remove the solvent. The residue was diluted with H ₂ O (500 ml) and was extracted using EtOAc (800 ml; 400 ml * 2). The combined organic layers were washed with aqueous NaCl (400 ml), dried, filtered and concentrated under reduced pressure to give the desired product. The compound iodomethyl (E)-octadec-9-en-1-yl carbonate (29 g, 64.10 mmol, yield: 77.13%) was obtained as a light yellow oil and used in the next step without further purification.

[00215] Step 7C: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl ((E)-octadec-9-en-1-yl) carbonate

[00216] A mixture of (4R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-7-one hydrochloride (8 g, 21.17 mmol, 1 eq.) and K ₂ CO ₃ (8.78 g, 63.52 mmol, 3 eq.) in H ₂ O (200 ml ) was stirred for 0.5 h at 25°C. Tetrabutylammonium sulfate (12.30 g, 21.17 mmol, 12.18 ml, 1 eq.) in DCM (200 ml) was added to the mixture and stirred for 0.5 h at 25°C. To this mixture was added iodomethyl (E)-octadec-9-en-1-yl carbonate (14.37 g, 31.76 mmol, 1.5 eq) and stirred for 11 hours. The reaction mixture was then concentrated under reduced pressure to remove DCM. The residue was diluted with H ₂ O (300 ml) and was extracted using EtOAc (300 ml * 3). The combined organic layers were washed with aqueous NaCl (300 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue. This crude product was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=10/1-3:1) to give the desired product. Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl [(E)-octadec-9-enyl] carbonate (8.08 g, 12.09 mmol, yield: 57.12%) was obtained as a colorless oil. M+H ⁺ = 666.5 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 7.

[00217] Example 8: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl (E)-octadec-9-enoate

[00218] DMF (N,N-dimethylformamide) (264 .03 mg, 3.61 mmol, 277.93 µl, 0.195 eq.) and oxalic acid dichloride (8.46 g, 66.69 mmol, 5.84 ml, 3.6 eq.) at 25°C in atmosphere _N2 . This mixture was stirred for 30 min at 25°C, then concentrated under reduced pressure. DCM (100 ml) was added to the residue. To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2- e]isoquinolin-7-one (7 g, 18.53 mmol, 1 eq, HCl) and DCM (100 mL) TEA (3.75 g, 37.05 mmol, 5.16 mL, 2 eq) was added , then to this mixture was added previously obtained mixture in portions at 25°C in an atmosphere of N ₂ . The resulting mixture was stirred for 12 hours at 25°C. The residue was concentrated in vacuo to remove DCM, then dissolved in saturated NaHCO ₃ solution (200 ml). The aqueous phase was extracted with ethyl acetate (100 ml * 2). The combined organic phases were dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=40/1-1/1). Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl](E)-octadec-9-enoate (5.36 g, 8.11 mmol, yield: 43.79%) was obtained as a yellow oil. M+H ⁺ = 606.2 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 8.

[00219] Example 9: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl (E)-octadec-9-enoate

[00220] To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2 -e]isoquinoline-4a,9-diol (10 g, 26.60 mmol, 1 eq, HCl) and H ₂ O (100 ml) were added in one go K ₂ CO ₃ (11.03 g, 79.81 mmol, 3 equiv.) at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. Tetrabutylammonium sulfate (30.91 g, 26.60 mmol, 30.61 ml, 50% solution, 1 eq.) and DCM (100 ml) were mixed, and then this mixture was added to the previously obtained mixture. Iodomethyl (E)-octadec-9-enoate (16.86 g, 39.91 mmol, 1.5 eq) prepared according to the procedure described in Example 41B was added and the mixture was stirred for 12 hours. The mixture was diluted with H ₂ O (100 ml), the organic phase was collected, then the aqueous phase was extracted with ethyl acetate (300 ml * 3), the organic phase was washed with brine (300 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=20/1-1/1). It was then further purified by preparative HPLC using MeOH as the solvent, standard reverse phase chromatography as the separation technique, and TFA as the separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (400 ml * 3). The combined organic phases were washed with brine (500 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl (E)-octadec-9-enoate (7.16 g, 11.15 mmol, yield: 41.91%) was obtained as a yellow oil. M+H ⁺ = 634.4 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 9.

[00221] Example 10: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl decyl carbonate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 1.31 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 10. Briefly, to a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-7-one (1.9 g, 5.57 mmol, 1 eq.) and DCM (15 ml) cooled to -10°C, TEA (1.69 g, 16. 70 mmol, 2.32 ml, 2 eq.) and decyl carbonochloridate (2.46 g, 11.13 mmol, 2 eq.). This mixture was stirred for 5 hours at 25°C. The reaction mixture was then concentrated under reduced pressure to leave a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 7/3 to 0:1) Compound [(4aS,7aR,12bS)-3-(cyclopropylmethyl )-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl decyl carbonate] (purity 97.43%) was obtained as a yellow oil with a yield of 43.63%.

[00222] Example 11: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl dodecyl carbonate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 11. Briefly, to a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-7-one (2.5 g, 6.62 mmol, 1 eq., HCl) and TEA (1.34 g, 13.23 mmol, 1.84 ml, 2 eq.) in DCM (15 ml) was added dodecyl carbonochloridate (1.56 g, 6.29 mmol, 0.95 eq.). This mixture was stirred for 1 hour at -10° C., then heated to 25° C. and stirred for 4 hours. The reaction mixture was then concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 7/3 to 0:1). This crude product was purified by preparative HPLC (column: Gemini 200*30 10 μm; mobile phase - [water (10 mm NH ₄ HCO ₃ )-CAN]; B% 70-100%, 12 min) Compound [(4aS,7aR ,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e] isoquinolin-9-yl dodecyl carbonate] (purity 99.497%) was obtained as a white solid (1.5 g, yield: 40.74%).

[00223] Example 12: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl stearate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.2 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 12.

[00224] Example 13: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl (Z)-docos-13-enoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.3 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 13.

[00225] Example 14: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl docosanoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 14. Briefly, to a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinoline-4a,9-diol (2 g, 5.32 mmol, 1 eq, HCl) and DCM (10 mL) TEA (1.62 g, 15.96 mmol, 2.22 mL, 3 eq.) and docosanoyl chloride (3.82 g, 10.64 mmol, 2 eq.) in one go at 25° C. under N ₂ . This mixture was stirred for 12 h at 25°C. The reaction mixture was then concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 5/1 to 1:1) The residue was purified by preparative HPLC using TFA (column: Phenomenx luna (2) C18 250*50 10 µm; mobile phase: [water (0.1% TFA)-CAN]; B%: 65-95%, 20 min]). Compound [(4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl docosanoate] (purity 97.01%) was obtained as a white solid (1.5 g, yield: 41.31%).

[00226] Example 15: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl (E)-octadec-9-enoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.8 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 15. Briefly, to a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinoline-4a,9-diol (2 g, 5.32 mmol, 1 eq, HCl) and DCM (30 mL) TEA (1.08 g, 10.64 mmol, 1.48 mL, 2 eq.) and (E)-octadec-9-enoyl chloride (1.92, 6.38 mmol, 1.2 eq.). This mixture was stirred for 12 hours at 15°C. Then the reaction mixture was mixed with H ₂ O (80 ml) and was extracted with DCM (80 ml x 3). The combined organic phases were washed with saturated NaHCO ₃ (60 ml x 2) and brine (60 ml x 3), dried using anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 10/1 to 1:1). The resulting compound was purified again using a preparative HPLC column: Phenomenex luna C18, 250 x 50 mm x 10 µm, mobile phase: [water (0.1% TFA)-CAN]; B: 60-90%, 20 min). After preparative HPLC, the mixture was concentrated under reduced pressure. The aqueous phase was mixed with NaHCO ₃ to obtain pH 8, then the aqueous phase was extracted with ethyl acetate (30 ml x 4). The combined organic phases were washed with brine (20 ml x 1), dried using anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo. Compound [(4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl (E)-octadec-9-enoate] (purity 95%) was obtained as a yellow oil (1.8 g, yield: 29.57%).

[00227] Example 16: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl eicosanoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.1 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 16. Briefly, to a solution of eicosanoic acid (5 g, 16.00 mmol, 5.92 ml, 1 eq.) in DCM (50 ml) was added DMF (116.93 mg, 1.6 mmol, 123.09 µl, 0.1 eq.), cooled to 0°C, (C°C l) ₂ (2.34 g, 18.40 mmol, 1.61 ml, 1.15 eq.), TEA (4.86 g , 48.80 mmol, 6.68 ml, 3 eq.) and (3R, 4aS, 7aS, 12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro- 1H-4,12-methanobenzofuro[3,2-e]isoquinoline-4a,9-diol (3.01 g, 8.00 mmol, 0.5 eq., HCl). This mixture was stirred for 12 hours at 25°C. Then the reaction mixture was extracted with H ₂ O (80 ml x 1) and DCM (80 ml x 2). The combined organic phases were washed with brine (60 ml x 3), dried using anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo. The compound was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 10/1 to 1:1). Compound [4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3 ,2-e]isoquinolin-9-yl eicosanoate] (purity 100%) was obtained as a white solid (1.1 g, yield: 10.84%).

[00228] Example 17: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl octyl carbonate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs and was obtained as an oil. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 17.

[00229] Example 18: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl decyl carbonate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs and was obtained as an oil. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 18.

[00230] Example 19: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl hexadecyl carbonate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.8 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 19. Briefly, to a solution of (3R, 4aS, 7aS, 12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinoline-4a,9-diol (5 g, 13.30 mmol, 1 eq.) in DCM (50 ml) cooled to -10° C. TEA (4.04 g, 39.91 mmol , 5.55 ml, 3 eq.) and hexadecyl carbonochloridate (8.11 g, 26.60 mmol, 2 eq.). Then this mixture was stirred for 5 hours at 25°C in an atmosphere of N ₂ . Then the reaction mixture was extracted with H ₂ O (80 ml x 1) and DCM (80 ml x 2). The combined organic phase was washed with brine (60 ml x 3), dried using anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue and compound were purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 10/1 to 1:1). The residue was purified by preparative HPLC using TFA (column - Phenomenex lune C18 250 x 50 mm x 10 μm, mobile phase - [water (0.1% TFA)-CAN]; B% 65-95%, 20 min). The pH was adjusted to 8 by adding NaHCO ₃ , [and then extracted with EtOAc (ethyl acetate) (20 ml x 3). The organic layer was evaporated under reduced pressure to give the final product. Compound [4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3 ,2-e]isoquinolin-9-yl hexadecyl carbonate] (purity 99.723%) was obtained as a white solid (1.8 g, yield: 12.33%).

[00231] Example 20: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl (9Z,12Z,15Z)-octadeca-9,12,15-trienoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.4 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 20.

[00232] Example 21: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl hexadecyl carbonate

[00233] The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 2.15 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 21. Briefly, to a mixture of (3R, 4aS, 7aR, 12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-7-one (2 g, 5.29 mmol, 1 eq, HCl) and DCM (20 ml) cooled to -10° C. TEA (1.61 g, 15. 88 mmol, 2.21 ml, 3 eq.) and hexadecyl carbonochloridate (3.23 g, 10.59 mmol, 2 eq.). Then this mixture was stirred for 5 hours at 25°C in an atmosphere of N ₂ . Then the reaction mixture was extracted with H ₂ O (80 ml x 1) and DCM (80 ml x 2). The residue was purified by preparative HPLC using TFA (column - Phenomenex luna (2) C18 250 x 50 mm x 10 µm, mobile phase - [water (0.1% TFA)-CAN]; B% 60-90%, 20 min ). The pH was adjusted to 8 by adding NaHCO ₃ and then extracted with EtOAc (ethyl acetate) (20 ml x 3). The organic layer was evaporated under reduced pressure to give the final product. Compound [(4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[ 3,2-e]isoquinolin-9-yl hexadecyl carbonate] (purity 97.669%) was obtained as a white solid (2.15 g, yield: 65.06%).

[00234] Example 22: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl (Z)-docos-13-enoate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 2.17 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 22.

[00235] Example 23: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl octyl carbonate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 1.33 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 23.

[00236] Example 24: Step 24A: Synthesis of chloromethyl dodecyl carbonate

To a mixture of dodecan-1-ol (30 g, 161.00 mmol, 1 eq.) and DCM (300 ml) was added TEA (32.58 g, 322.00 mmol, 44.82 ml, 2 eq. ) and chloroformic acid chloromethyl ether (41.52 g, 322.00 mmol, 28.63 ml, 2 eq.) at 0°C under N ₂ atmosphere. This mixture was heated to 25°C and stirred for 12 hours. Then the reaction mixture was quenched by adding water (200 ml) at 25°C and then extracted using DCM (100 ml; 50 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–80:1). The compound chloromethyl dodecyl carbonate (10.3 g, 36.94 mmol, yield: 22.95%) was obtained as a colorless oil.

[00237] Step 24B: Synthesis of iodomethyl dodecyl carbonate

[00238] NaHCO ₃ (3.62 g, 43.04 mmol, 1.67 ml, 1 .2 eq.) and NaI (6.45 g, 43.04 mmol, 1.2 eq.) at 25° C. under N ₂ atmosphere. This mixture was stirred in the dark for 12 hours at 25°C. The reaction mixture was then filtered to remove insoluble matter and concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate (50 ml), and the organic layer was washed with water (60 ml; 30 ml * 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue. The compound dodecyl iodomethyl carbonate (12.6 g, crude) was obtained as a light red oil. The crude dodecyl iodomethyl carbonate was used in the next step without further purification.

[00239] Step 24C: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecyl carbonate

To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e] isoquinoline-4a,9-diol (4 g, 10.64 mmol, 1 eq., HCl) and H ₂ O (20 ml) was added K ₂ CO ₃ (4.41 g, 31.92 mmol, 3 eq.) , and this mixture was stirred for 30 min at 20°C. Tetrabutylammonium sulfate (12.37 g, 10.64 mmol, 12.24 ml, 1 eq.) and DCM (20 ml) were added to the mixture, and the resulting mixture was stirred for 10 min at 20°C. Dodecyl iodomethyl carbonate (9.46 g, 25.54 mmol, 2.4 eq.) was added to the mixture in one go at 20° C. under N ₂ atmosphere. The resulting mixture was stirred for 12 hours at 20°C. Then the reaction mixture was diluted with water (20 ml) and extracted with DCM (20 ml; 10 ml * 2). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1:0–10:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl dodecyl carbonate (2.92 g, 5.00 mmol, yield: 47.02%) was obtained as a colorless oil. M+H ⁺ = 582.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 24.

[00240] Example 25: Step 25A: Synthesis of chloromethyl tetradecyl carbonate

To a mixture of tetradecan-1-ol (30 g, 139.93 mmol, 1 eq.) and DCM (300 ml) was added TEA (28.32 g, 279.87 mmol, 38.95 ml, 2 eq. ) and chloroformic acid chloromethyl ether (36.09 g, 279.87 mmol, 24.89 ml, 2 eq.) at 0° C. under N ₂ , then heated to 25° C. for 12 h. The reaction mixture was quenched by adding water (200 ml) at 25°C and then extracted using DCM (100 ml; 50 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–80:1). The compound chloromethyl tetradecyl carbonate (11 g, 35.85 mmol, yield: 25.62%) was obtained as a colorless oil.

[00241] Example 25B: Synthesis of iodomethyl tetradecyl carbonate

To a mixture of chloromethyl tetradecyl carbonate (11.1 g, 36.17 mmol, 1 eq.) and acetone (100 ml) was added NaHCO ₃ (3.04 g, 36.17 mmol, 1.41 ml, 1 eq. .) and NaI (5.42 g, 36.17 mmol, 1 eq.) at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. The reaction mixture was then concentrated under reduced pressure to remove the solvent. The residue was diluted with ethyl acetate (40 ml) and washed with water (40 ml; 20 ml * 2). The organic layers were dried, filtered and concentrated under reduced pressure to give the crude iodomethyl tetradecyl carbonate (13.1 g, 32.89 mmol, yield: 90.92%) as a light red oil, which was used in the next step without additional cleaning.

[00242] Step 25C: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl tetradecyl carbonate

To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e] isoquinoline-4a,9-diol (3 g, 7.98 mmol, 1 eq., HCl) and iodomethyl tetradecyl carbonate (7.63 g, 19.15 mmol, 2.4 eq.) in H ₂ O (30 ml ) was added K ₂ CO ₃ (3.31 g, 23.94 mmol, 3 eq.) and the mixture was stirred for 0.5 h at 15°C. After 0.5 h, tetrabutylammonium sulfate (4.64 g, 7.98 mmol, 4.59 ml, 1 eq.) and DCM (30 ml) were added to the mixture, and the mixture was stirred for 10 min at 15°C. After 10 minutes, iodomethyl tetradecyl carbonate (7.63 g, 19.15 mmol, 2.4 eq.) was added to the mixture in one go at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. The residue was diluted with water (10 ml) and extracted using DCM (20 ml; 10 ml * 2). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1:0–10:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl tetradecyl carbonate (2.0 g, 3.25 mmol, yield: 40.76%) was obtained as a colorless oil. M+H ⁺ = 610.5 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 25.

[00243] Example 26: Step 26A: Synthesis of chloromethyl (E)-octadec-9-enoate

[00244] NaHCO _{3 (} 2 .38 g, 28.32 mmol, 1.10 ml, 4 eq.) and tetrabutylammonium sulfate (822.29 mg, 708.06 µmol, 50% solution, 0.1 eq.) at 25°C under N ₂ , then this mixture was cooled to 0°C. To the mixture was added chloro(chlorosulfonyloxy)methane (1.17 g, 7.08 mmol, 1 eq.) in DCM (10 ml) at 0°C. This mixture was heated to 25°C and stirred for 18 hours. Then the reaction mixture was extracted using DCM (30 ml; 15 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to give a residue. The compound chloromethyl (E)-octadec-9-enoate (1.97 g, 5.95 mmol, yield: 84.07%) was obtained as a white solid, which was used without purification in the next step.

[00245] Step 26B: Synthesis of iodomethyl (E)-octadec-9-enoate

[00246] NaHCO ₃ (4.42 g, 52, 58 mmol, 2.04 ml, 1.2 eq.) and NaI (7.88 g, 52.58 mmol, 1.2 eq.) at 25°C under N ₂ atmosphere. This mixture was stirred in the dark for 12 hours at 25°C. The reaction mixture was then filtered to remove insoluble matter and concentrated under reduced pressure. The resulting residue was dissolved in ethyl acetate (100 ml) and the organic layer was washed with brine (100 ml), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to obtain a residue. The compound iodomethyl (E)-octadec-9-enoate (18.6 g, crude) was obtained as a brown oil, which was used without purification in the next step.

[00247] Step 26C: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl (E)-octadec-9-enoate

[00248] To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3 ,2-e]isoquinolin-7-one (10.09 g, 26.71 mmol, 1 eq., HCl) and H ₂ O (100 ml) were added in one go K ₂ CO ₃ (11.07 g, 80 ,12 mmol, 3 eq.) at 25°C under N ₂ atmosphere. This mixture was stirred for 30 min at 25°C. Tetrabutylammonium sulfate (15.01 g, 12.92 mmol, 50% solution, 4.84e-1 eq.) and DCM (100 ml) were mixed, and then this mixture was added to the previously obtained mixture. Iodomethyl (E)-octadec-9-enoate (16.92 g, 40.06 mmol, 1.5 eq.) was added and the mixture was stirred for 12 hours. The mixture was diluted with H ₂ O (800 ml), the organic layer was collected, then extracted with ethyl acetate (300 ml * 3). The entire organic phase was washed with brine (300 ml), dried over Na ₂ SO ₄ , filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (petroleum ether/ethyl acetate=20/1-1/1). The residue was further purified by preparative HPLC using MeOH as solvent, standard reverse phase chromatography separation technique, and TFA separation system. The pH was adjusted to approximately 8 by adding NaHCO ₃ , the aqueous phase was extracted with ethyl acetate (400 ml * 3). The combined organic phases were washed with brine (500 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl (E)-octadec-9-enoate (10.20 g, 15.85 mmol, yield: 59.35%) was obtained as a yellow oil. M+H ⁺ = 636.4 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 26.

[00249] Example 27: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl tetradecyl carbonate

To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2- e]isoquinolin-7-one (4.5 g, 11.91 mmol, 1 eq., HCl) and H ₂ O (30 ml) were added in one go K ₂ CO ₃ (4.94 g, 35.73 mmol , 3 equiv.) at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. To the mixture were added tetrabutylammonium sulfate (13.84 g, 11.91 mmol, 13.70 ml, 50% solution, 1 eq.) and DCM (30 ml) at 25° C. under N ₂ in one portion. This mixture was stirred for 10 min at 25°C. Iodomethyl tetradecyl carbonate (11.38 g, 28.58 mmol, 2.4 eq.) was added to the mixture in one go at 25° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 25°C. Then the reaction mixture was extracted using DCM (30 ml; 15 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–5:1). Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl tetradecyl carbonate (2.8 g, 4.53 mmol, yield: 38.05%, purity: 99%) was obtained as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 27.

[00250] Example 28: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl eicosyl carbonate

To a mixture of eicosyl (4-nitrophenyl) carbonate (9.87 g, 21.28 mmol, 4 eq.) and DCM (40 ml) was added TEA (538.40 mg, 5.32 mmol, 740.58 μl , 1 eq.) and (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinoline-4a,9-diol (2 g, 5.32 mmol, 1 eq., HCl) at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. Then the reaction mixture was extracted using H ₂ O (20 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by preparative HPLC. Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]eicosyl carbonate (1.6 g, 2.35 mmol, yield: 44.25%) was obtained as a white solid. ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 28.

[00251] Example 29: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecyl carbonate

To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2- e]isoquinolin-7-one (4.5 g, 11.91 mmol, 1 eq., HCl) and H ₂ O (30 ml) were added in one go K ₂ CO ₃ (4.94 g, 35.73 mmol , 3 equiv.) at 25°C in an atmosphere of N ₂ . This mixture was stirred for 30 min at 25°C. To the mixture were added tetrabutylammonium sulfate (13.84 g, 11.91 mmol, 13.70 ml, 50% solution, 1 eq.) and DCM (30 ml) at 25° C. under N ₂ in one portion. This mixture was stirred for 10 min at 25°C. Dodecyl iodomethyl carbonate (10.58 g, 28.58 mmol, 2.4 eq.) was added to the mixture in one go at 25° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 25°C. Then the reaction mixture was extracted using DCM (30 ml; 15 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–5:1). Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl dodecyl carbonate (3.1 g, 5.26 mmol, yield: 44.19%, purity: 99.1%) was obtained as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 29.

[00252] Example 30: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl tridecyl carbonate

To a mixture of (4-nitrophenyl) tridecyl carbonate (5.83 g, 15.96 mmol, 2 eq.) and DCM (50 ml) was added TEA (2.42 g, 23.94 mmol, 3.33 ml , 3 eq.) and (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinoline-4a,9-diol (3 g, 7.98 mmol, 1 eq., HCl) at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. The reaction mixture was then concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–3:1). [(3R,4aS,7aS,12bS)-3-(Cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2 -e]isoquinolin-9-yl]tridecyl carbonate (2.3 g, 4.07 mmol, yield: 50.94%) was obtained as a colorless oil. ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. thirty.

[00253] Example 31: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl tetradecyl carbonate

To a solution of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e] isoquinoline-4a,9-diol (3 g, 7.98 mmol, 1 eq., HCl) in DCM (20 ml) was added TEA (1.62 g, 15.96 mmol, 2.22 ml, 2 eq.) and chloroformic acid tetradecyl ester (2.21 g, 7.98 mmol, 1 eq.). This mixture was stirred for 12 hours at 15°C. The mixture was then concentrated under reduced pressure. The residue was mixed with H ₂ O (80 ml) and was extracted using DCM (80 ml * 3). The combined organic phases were washed with saturated NaHCO ₃ solution (60 ml*2) and brine (60 ml*3), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=4/1–0:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]tetradecyl carbonate (2 g, 3.41 mmol, yield: 42.79%) was obtained as a colorless oil. M+H ⁺ = 580.4 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 31.

[00254] Example 32: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl pentadecyl carbonate

To a mixture of (4-nitrophenyl)pentadecyl carbonate (6.28 g, 15.96 mmol, 2 eq.) and DCM (30 ml) was added TEA (2.42 g, 23.94 mmol, 3.33 ml , 3 eq.) and [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3 ,2-e]isoquinoline-4a,9-diol (3 g, 7.98 mmol, 1 eq., HCl) at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–2:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]pentadecyl carbonate (2.6 g, 1.80 mmol, yield: 22.49%) was obtained as a white solid. M+H ⁺ = 594.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 32.

[00255] Example 33: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl octadecyl carbonate

To a solution of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e] isoquinoline-4a,9-diol (2 g, 5.32 mmol, 1 eq, HCl) in DCM (30 ml) was added TEA (1.62 g, 15.96 mmol, 2.22 ml, 3 eq) and (4-nitrophenyl) octadecyl carbonate (3.48 g, 7.98 mmol, 1.5 eq.). This mixture was stirred for 12 hours at 15°C. The reaction mixture was then concentrated under reduced pressure. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=10/1–1:1) and then by preparative HPLC. Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]octadecyl carbonate (0.8 g, 1.22 mmol, yield: 22.93%) was obtained as a yellow oil. M+H ⁺ = 636.5 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 33.

[00256] Example 34: Step 34A: Synthesis of chloromethyl hexadecyl carbonate

To a mixture of hexadecan-1-ol (30 g, 123.74 mmol, 1 eq.) and DCM (200 ml) was added TEA (25.04 g, 247.48 mmol, 34.45 ml, 2 eq. ) and chloroformic acid chloromethyl ether (31.91 g, 247.48 mmol, 22.01 ml, 2 eq.) at 0°C under N ₂ atmosphere. This mixture was heated to 20°C and stirred for 12 hours. Then the reaction mixture was quenched by adding water (50 ml) at 20°C and then extracted using DCM (100; ml 50 ml * 2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1/0–80:1). The compound chloromethyl hexadecyl carbonate (18 g, 53.74 mmol, yield: 43.43%) was obtained as a white solid.

[00257] Step 34B: Synthesis of iodomethyl hexadecyl carbonate

[00258] NaHCO ₃ (2.41 g, 28.66 mmol, 1.11 ml, 1 ,2 eq.) and NaI (4.30 g, 28.66 mmol, 1.2 eq.) at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. The reaction mixture was then concentrated under reduced pressure to remove the solvent. The residue was diluted with ethyl acetate (20 ml) and washed with water (20 ml; 10 ml * 2). The organic layers were dried, filtered and concentrated under reduced pressure to give the crude product hexadecyl iodomethyl carbonate (9 g, crude) as a light red solid, which was used in the next step without further purification.

Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro [3,2-e]isoquinolin-9-yl)oxy)methyl hexadecyl carbonate

[00259] To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2 -e]isoquinoline-4a,9-diol (2.75 g, 7.32 mmol, 1 eq., HCl) and hexadecyl iodomethyl carbonate (7.49 g, 17.56 mmol, 2.4 eq.) in H ₂ O (25 ml) K ₂ CO ₃ (3.03 g, 21.95 mmol, 3 eq.) was added and stirred for 0.5 h at 15°C. After 30 minutes, tetrabutylammonium sulfate (4.25 g, 7.32 mmol, 4.21 ml, 1 eq.) and DCM (25 ml) were added to the mixture, and the mixture was stirred for over 10 minutes. After 10 minutes, hexadecyl iodomethyl carbonate (7.49 g, 17.56 mmol, 2.4 eq.) was added to the mixture in one go at 15° C. under N ₂ atmosphere. This mixture was stirred for 12 hours at 15°C. The residue was diluted with water (10 ml) and extracted using DCM (20 ml; 10 ml * 2). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate=1:0–10:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl hexadecyl carbonate (2.0 g, 3.10 mmol, yield: 42.38%) was obtained as a colorless oil. M+H ⁺ = 638.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 34.

[00260] Example 35: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl decyl carbonate

[00261] (((4aS,7aS,12bS)-3-(Cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12 -methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl decyl carbonate was prepared analogously to the compound of Example 5. To a mixture of (3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2, 4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinoline-4a,9-diol and H ₂ O were added in one go K ₂ CO ₃ at 25°C in N ₂ atmosphere. This mixture was stirred for 30 min at 25°C. Then tetrabutylammonium sulfate in DCM was added in one go at 25°C. Then, iodomethyl decyl carbonate was added to the reaction mixture, and the mixture was stirred at 25° C. until the reaction was completed. The reaction mixture was then further worked up and the desired product was isolated by chromatography as described in Example 5. ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 35.

[00262] Example 36: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl oleate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 36.

[00263] Example 37: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl (9Z,12Z)-octadeca-9,12-dienoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.3 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 37.

[00264] Example 38: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3,3-dimethylbutanoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.2 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 38.

[00265] Example 39: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3-cyclopentylpropanoate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs. 1.18 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 39.

[00266] Example 40: Synthesis of (4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl tert-butyl carbamate

The title compound was synthesized according to General Scheme 1 for the synthesis of nalmefene prodrugs and obtained as a solid. 3.0 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 40.

[00267] Example 41: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl oleate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 1.5 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 41.

[00268] Example 42: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3,3-dimethylbutanoate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 1.3 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 42.

[00269] Example 43: Synthesis of (4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4 ,12-methanobenzofuro[3,2-e]isoquinolin-9-yl 3-cyclopentylpropanoate

The title compound was synthesized according to General Scheme 2 for the synthesis of naltrexone prodrugs. 1.3 g, ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 43.

[00270] Example 44: Step 44A: Synthesis of chloromethyl dodecanoate

_{NaHCO 3} (33.55 g, 399.37 mmol, 15.53 ml, 4 eq.) and tetrabutylammonium sulfate (11.60 g, 9.98 mmol, 11.49 ml, 50% purity, 0.1 eq.) in one go at 25° C. under N ₂ , then this mixture was cooled to 0°C. The reagent chloro(chlorosulfonyloxy)methane (16.47 g, 99.84 mmol, 1 eq.) in DCM (20 mL) was added to the mixture in one go at 0°C. The resulting mixture was heated to 25°C and stirred for 18 hours. The reaction mixture was then extracted with 50 ml DCM (25 ml*2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1/0 to 80:1). The compound chloromethyl dodecanoate (10.8 g, 43.41 mmol, yield: 43.48%) was obtained as a colorless oil.

[00271] Step 44B: Synthesis of iodomethyl dodecanoate

A mixture of chloromethyl dodecanoate (9 g, 36.18 mmol, 1 eq.) and acetone (80 ml) was degassed and purged with nitrogen 3 times, and then NaHCO ₃ (3.04 g, 36.18 mmol, 1 41 ml, 1 eq.) and NaI (5.42 g, 36.18 mmol, 1 eq.), the resulting mixture was stirred in the dark for 12 h at 15° C. under N ₂ atmosphere. The reaction mixture was then filtered and concentrated under reduced pressure to remove the solvent. The residue was diluted with H ₂ O (50 ml) and was extracted with EtOAc (ethyl acetate) (120 ml). The combined organic layers were washed with 100 ml H ₂ O (50 ml*2), dried, filtered and concentrated under reduced pressure to give a residue. The compound iodomethyl dodecanoate (9 g, crude) was obtained as a yellow liquid and was used in the next step without further purification.

[00272] Step 44C: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecanoate

(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinoline mixture -4a,9-diol (3.5 g, 10.31 mmol, 1 eq.), K ₂ CO ₃ (4.28 g, 30.93 mmol, 3 eq.) in H ₂ O (40 ml) was stirred for 30 min at 15°C, then tetrabutylammonium sulfate (5.99 g, 10.31 mmol, 5.93 ml, 1 eq.) and DCM (20 ml) were added to the mixture, and a solution of iodomethyl dodecanoate was added to the resulting mixture (8.42 g, 24.75 mmol, 2.4 eq.) in DCM (20 ml) and degassed and purged with nitrogen 3 times, and then this mixture was stirred for 11.5 h at 15° C. under N ₂ . Then the reaction mixture was diluted with H ₂ O (20 ml) and was extracted with DCM (20 ml). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1:0 to 20:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl dodecanoate (1.51 g, 2.70 mmol, yield: 26.14%) was obtained as a colorless oil. M+H ⁺ = 552.5 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 44.

[00273] Example 45: Step 45A Synthesis of chloromethyl tetradecanoate

[00274] To a mixture of tetradecanoic acid (20 g, 87.58 mmol, 1 eq.) and H ₂ O (80 ml) was added NaHCO ₃ (29.43 g, 350.31 mmol, 13.62 ml, 4 eq. ) and tetrabutylammonium sulfate (10.18 g, 8.76 mmol, 10.08 ml, 50% solution, 0.1 eq.) and DCM (60 ml) under N ₂ atmosphere. This mixture was cooled to 0°C. The reagent chloro(chlorosulfonyloxy)methane (14.45 g, 87.58 mmol, 1 eq.) in DCM (20 ml) was added to the mixture in one go at 0° C. under N ₂ atmosphere. The resulting mixture was heated to 25°C and stirred for 18 hours. The reaction mixture was then extracted with 50 ml DCM (25 ml*2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1/0 to 80:1). The compound chloromethyl tetradecanoate (15.5 g, 55.99 mmol, yield: 63.93%) was obtained as a colorless oil.

[00275] Step 45B: Synthesis of iodomethyl tetradecanoate

A mixture of chloromethyl tetradecanoate (8 g, 28.90 mmol, 1 eq.) and acetone (70 ml) was degassed and purged with nitrogen 3 times, and then NaHCO ₃ (2.43 g, 28.90 mmol, 1 , 12 ml, 1 eq.) and NaI (4.33 g, 28.90 mmol, 1 eq.), the resulting mixture was stirred in the dark for 12 h at 15° C. under N ₂ atmosphere. The reaction mixture was then filtered and concentrated under reduced pressure to remove the solvent. The residue was diluted with H ₂ O (50 ml) and was extracted with EtOAc (ethyl acetate) (120 ml). The combined organic layers were washed with 100 ml H ₂ O (50 ml * 2), dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure to leave the crude product, which was used in the next step without further purification. The compound iodomethyl tetradecanoate (9 g, crude) was obtained as a yellow solid.

[00276] Step 45C: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl tetradecanoate

(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinoline mixture -4a,9-diol (3.5 g, 9.31 mmol, 1 eq., HCl), K ₂ CO ₃ (3.86 g, 27.93 mmol, 3 eq.) in H ₂ O (30 ml ) was stirred for 30 min at 15°C, then tetrabutylammonium sulfate (5.41 g, 9.31 mmol, 5.36 ml, 1 eq.) and DCM (15 ml) were added to the mixture, and a solution of iodomethyl tetradecanoate (8.23 g, 22.35 mmol, 2.4 eq.) in DCM (15 ml) and degassed and purged with nitrogen 3 times, and then this mixture was stirred for 11.5 h at 15°C in N ₂ atmosphere. Then the reaction mixture was diluted with H ₂ O (20 ml) and was extracted with DCM (20 ml). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1:0 to 20:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl tetradecanoate (1.5 g, 2.54 mmol, yield: 27.26%) was obtained as a colorless oil. M+H ⁺ = 580.5 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 45.

[00277] Example 46: Step 46A: Synthesis of chloromethyl hexadecanoate

_[ 00278] NaHCO ₃ (26.21 g, 311, 98 mmol, 12.13 ml, 4 eq.) and tetrabutylammonium sulfate (9.06 g, 7.80 mmol, 8.97 ml, 50% pure, 0.1 eq.) in a single dose at 25°C in the atmosphere N ₂ and then this mixture was cooled to 0°C. The reagent chloro(chlorosulfonyloxy)methane (12.87 g, 78.00 mmol, 1 eq.) in DCM (20 mL) was added to the mixture in one go at 0°C. The resulting mixture was heated to 25°C and stirred for 18 hours. The reaction mixture was then extracted with 50 ml DCM (25 ml*2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1/0 to 80:1). The compound chloromethyl hexadecanoate (17.6 g, 57.72 mmol, yield: 74.01%) was obtained as a white solid.

[00279] Step 46B: Synthesis of iodomethyl hexadecanoate

A mixture of chloromethyl hexadecanoate (3 g, 9.84 mmol, 1 eq.) and acetone (30 ml) was degassed and purged with nitrogen 3 times at 15°C in the dark, and then NaHCO ₃ (826.58 mg, 9 .84 mmol, 382.68 μl, 1 eq.) and NaI (1.47 g, 9.84 mmol, 1 eq.), and stirred in the dark for 12 h at 15° C. under N ₂ atmosphere. The reaction mixture was then filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was diluted with H ₂ O (20 ml) and was extracted with EtOAc (ethyl acetate) (60 ml). The combined organic layers were washed with 40 ml H ₂ O (20 ml*2), dried, filtered and concentrated under reduced pressure to give a residue. The compound iodomethyl hexadecanoate (3.5 g, crude) was obtained as a yellow solid and was used in the next step without further purification.

[00280] Step 46C: Synthesis of (((4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecanoate

(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinoline mixture -4a,9-diol (1.8 g, 4.79 mmol, 1 eq., HCl), K ₂ CO ₃ (1.99 g, 14.37 mmol, 3 eq.) in H ₂ O (15 ml ) was stirred for 30 min, tetrabutylammonium sulfate (2.78 g, 4.79 mmol, 2.75 ml, 1 eq.) and DCM (7.5 ml) were added to the mixture, and a solution of iodomethyl hexadecanoate (4.56 g, 11.49 mmol, 2.4 eq.) in DCM (7.5 ml), and degassed and purged with nitrogen 3 times, and then this mixture was stirred for 11.5 h at 15°C in N ₂ atmosphere. Then the reaction mixture was diluted with H ₂ O (10 ml) and was extracted with DCM (10 ml). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1:0 to 20:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl hexadecanoate (1.6 g, yield: 24.35%) was obtained as a colorless oil. M+H ⁺ = 608.6 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 46.

[00281] Example 47: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecyl carbonate

To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2- e]isoquinolin-7-one (5.45 g, 14.42 mmol, 1 eq, HCl) and H ₂ O (30 ml) were added in one go at 25° C. under N ₂ K ₂ CO ₃ (5 .98 g, 43.27 mmol, 3 eq.). This mixture was stirred for 30 min at 25°C. Tetrabutylammonium sulfate (16.76 g, 14.42 mmol, 16.59 ml, 50% solution, 1 eq.) and DCM (30 ml) were added to the mixture at 25°C, and the mixture was stirred for 10 min at 25 °C Hexadecyl iodomethyl carbonate (14.76 g, 34.62 mmol, 2.4 eq.) was added to the mixture in one go at 25°C and the mixture was stirred for 12 h at 25°C. The reaction mixture was then extracted with 30 ml DCM (15 ml*2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1/0 to 5:1). Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl hexadecyl carbonate (4.68 g, 7.29 mmol, yield: 50.56%) was obtained as a colorless oil. M+H ⁺ = 640.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 47.

[00282] Example 48: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl dodecanoate

(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinoline mixture -4a,9-diol (3.5 g, 10.31 mmol, 1 eq.), K ₂ CO ₃ (4.28 g, 30.93 mmol, 3 eq.) in H ₂ O (40 ml) was stirred for 30 min at 15°C, then tetrabutylammonium sulfate (5.99 g, 10.31 mmol, 5.93 ml, 1 eq.) and DCM (20 ml) were added to the mixture, and a solution of iodomethyl dodecanoate was added to the resulting mixture (8.42 g, 24.75 mmol, 2.4 eq.) in DCM (20 ml) and degassed and purged with nitrogen 3 times, and then this mixture was stirred for 11.5 h at 15° C. under N ₂ . Then the reaction mixture was diluted with H ₂ O (20 ml) and was extracted with DCM (20 ml). The combined organic layers were dried, filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1:0 to 20:1). Compound [(3R,4aS,7aS,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-methylene-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl dodecanoate (1.51 g, 2.70 mmol, yield: 26.14%) was obtained as a colorless oil. M+H ⁺ = 554.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 48.

[00283] Example 49: Synthesis of (((4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H -4,12-methanobenzofuro[3,2-e]isoquinolin-9-yl)oxy)methyl hexadecanoate

To a mixture of (3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a,9-dihydroxy-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3,2- e]isoquinolin-7-one (5 g, 13.23 mmol, 1 eq, HCl) and H ₂ O (30 ml) were added in one go at 25° C. under N ₂ K ₂ CO ₃ (5.49 g, 39.70 mmol, 3 eq.). This mixture was stirred for 30 min at 25°C. Tetrabutylammonium sulfate (15.38 g, 13.23 mmol, 15.22 ml, 50% solution, 1 eq.) and DCM (30 ml) were added to the mixture in one go at 25° C. under N ₂ atmosphere. This mixture was stirred for 10 min at 25°C. Iodomethyl hexadecanoate (12.59 g, 31.76 mmol, 2.4 eq.) was added to the mixture in one go at 25° C. under N ₂ atmosphere. This mixture was stirred for 12 h at 25°C. The reaction mixture was then extracted with 30 ml DCM (15 ml*2). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated under reduced pressure. And the resulting residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate 1/0 to 5:1). Compound [(3R,4aS,7aR,12bS)-3-(cyclopropylmethyl)-4a-hydroxy-7-oxo-2,4,5,6,7a,13-hexahydro-1H-4,12-methanobenzofuro[3, 2-e]isoquinolin-9-yl]oxymethyl hexadecanoate (5.2 g, 6.14 mmol, yield: 46.40%) was obtained as a light yellow oil. M+H ⁺ = 610.3 (LC-MS). ¹ H NMR (400 MHz, CDCl ₃ ): see FIG. 49.

Examples 50-61. Compounds 50-61 shown in Table 2 can be prepared according to General Scheme 1 for the synthesis of nalmefene prodrugs or General Scheme 2 for the synthesis of naltrexone prodrugs using suitable starting materials.

II. Biological assessment

Example 1 Stability Assay in Plasma and Liver S9 Fraction

[00284] Determination of the stability of the test compounds in the plasma of rats, dogs, cynomolgus monkeys and humans was performed using HPLC-MS (high performance liquid chromatography/mass spectrometry). Rat Plasma Analysis: Five aliquots of 70 μl each (one aliquot for each time point) were incubated in a 96-well polypropylene plate. Test compounds (10 μM, final solvent concentration 1%) were incubated at 37°C. Five time points were analyzed (0, 15, 120, 480, and 1440 min). Canine, monkey and human plasma analysis: Test compounds (2 μM, final solvent concentration 1%) were also incubated at 37° C. and analyzed at five time points (0, 10, 30, 60 and 120 min). All incubations were performed in duplicate. Samples were analyzed using HPLC-MS. The amount of parent compound (in percent) remaining after incubation in plasma was determined. Data have previously been published for nalmefene dodecanoate and nalmefene palmitate (Gaekens et al., Journal of Controlled Release 232 (2016) 196-202). The results are shown in Table 4a-d.

Table 4a. Stability in Rat Plasma % remaining at each time point expressed in minutes Compound 0 min 15 minutes 120 min 480 min 1440 min 36 100 74.4 66.3 39.8 19.1 6 100 31.3 7.6 4 1.6 53 100 46.85 19.6 7.3 1.9 eleven 100 0.7 0.3 0.2 0 54 100 98.8 62.4 45.6 19.4 14 100 94 112 107 120 15 100 72 69.3 52 35 17 100 0 0 0 0 18 100 0.6 0.3 0.1 0 19 100 70.1 71.5 43.8 33.3 55 100 77.3 47.2 27.1 15.6 23 100 0 0 0 0 10 100 0 0 0 0 21 100 73.5 68.3 59.8 40.9 50 100 114.5 102.7 83.3 73.9 51 100 107.4 105.3 88.2 92.7 8 100 44.2 34.3 15.3 5.2 57 100 10.5 3.2 1.2 0 58 100 37 20 2.9 1.8 thirty 100 65.7 40.3 30.5 22.2 31 100 123.2 73.5 54.4 46.4 24 100 32 11.8 8.2 3 25 100 82.6 69.3 62.2 51.7 59 100 5.9 0.7 0.2 0 56 100 73.1 56.7 35.8 16.8 Aripiprazole lauroxyl (plasma breakdown control) 100 84 85.5 76.4 46 Enalapril maleate (salt) (control of plasma breakdown) 100 44 2.9 8.2 4.8

Table 4b. Stability in dog plasma % remaining at each time point expressed in minutes Compound 0 min 15 minutes 120 min 480 min 1440 min 6 100 89.1 96.9 107.2 101.2 15 100 82.5 80.6 90.1 86.1 24 100 94.7 104.5 115.7 116.2 Aripiprazole lauroxyl 100 103 81.3 78.6 74.3 paliperidone palmitate 100 91.2 88.4 95.9 93.1

Table 4c. Monkey Plasma Stability % remaining at each time point expressed in minutes Compound 0 min 15 minutes 120 min 480 min 1440 min 6 100 88.7 74.3 68.4 70.1 15 100 74.5 72.7 80.6 76.5 24 100 90.7 93.5 96.2 97.8 Aripiprazole lauroxyl 100 95.2 94.5 75.5 83.2 paliperidone palmitate 100 92 89.5 98.4 92.5

Table 4d. Stability in human plasma % remaining at each time point expressed in minutes Compound 0 min 15 minutes 120 min 480 min 1440 min 6 100 97.6 102 109.8 115.1 15 100 83.3 82.5 107.1 109.6 24 100 94 102.3 105.3 112.5 Aripiprazole lauroxyl 100 99.7 80 73.2 75.5 paliperidone palmitate 100 103.7 97.6 109.1 104.1

[00285] Determination of the stability of test compounds in the S9 fraction of the liver of dogs, cynomolgus monkeys and humans was performed using HPLC-MS. The stability of the test compound (2 μM, final concentration of DMSO - 0.1%, methanol - 1%) was evaluated in 50 μl of phosphate buffer containing 1.0 mg/ml of S9 protein isolated from each of the three species, and 5 mm D- sugar acid-1, 4-lactone. Samples were incubated for 60 minutes at 37°C and the remaining amount of the compound, expressed as %, was evaluated.

Table 4e. Stability in the S9 fraction of the liver % remaining after 60 minutes Compound Dog Monkey Human 6 17.1 0 5.3 15 80.9 76.7 59.8 24 3.6 3 3.5 Aripiprazole lauroxyl 66.1 56.6 47.8 paliperidone palmitate 56.4 57.4 47.8

Example 2 Opioid Receptor Binding Assay

[00286] A receptor binding assay was performed to evaluate the ability of compounds to inhibit binding to a radiolabeled ligand. First of all, the IC50 values of the selected compounds were determined for all three subtypes of opioid receptor subtypes (DOR, MOR and KOR), and these values were compared with the corresponding values of the parent molecule, namely: nalmefene. Based on the general experimental data, it can be concluded that derivatization to prodrugs significantly reduces the binding affinity for opioid receptors, in some cases by several orders of magnitude.

Equipment

Filter plates Unifilter-96 GF/C, Perkin Elmer (Cat#6005174)

96-well polypropylene conical plates, Agilent (Cat#5042-385)

Sealing film TopSeal-A, Perkin Elmer (Cat#6005250)

TopCount NXT HTS, (PerkinElmer)

MicroBeta2 (PerkinElmer)

Cell Collector C961961 (Perkin Elmer)

Reagents

Received stable cell lines, and using these cell lines received preparations of cell membranes.

³ H-diprenophrine (PerkinElmer, Cat: NET1121250UC, Lot: 2143599)

³ H-DAMGO (PerkinElmer, Cat: NET902250UC, Lot: 2139100)

³ H-DADLE (PerkinElmer, Cat: NET648250UC, Lot: 2060549)

Tris base (Sigma, Cat: T6066-1KG) was prepared as a 1 M stock solution and the pH was adjusted to 7.4.

0.5 M EDTA (ethylenediaminetetraacetic acid) (Invitrogen, Cat: 15575-038)

1 M MgCl ₂ (Sigma, Cat: M1028-100 ml)

PEI (Polyethyleneimine) (Sigma, Cat: P3143)

Microscint 20 c°C ktail (PerkinElmer, Cat: 6013329)

Naltrindole (Sigma, Cat: N115)

(±) trans-U-50488 (Sigma, Cat: D8040)

DAMGO (Sigma, Cat: E7384)

Analysis Buffer

Opioid receptor delta
Analysis Buffer Ultimate
concentration Tris-HCl 50 mM MgCl ₂ 10 mM EDTA 1 mM pH adjusted to 7.4
storage at 4°C

Opioid receptor kappa
Analysis Buffer ultimate
concentration Tris-HCl 50 mM pH adjusted to 7.4
storage at 4°C

Mu opioid receptor
Analysis Buffer ultimate
concentration Tris-HCl 50 mM MgCl ₂ 5 mM pH adjusted to 7.4
storage at 4°C

Wash Buffer

Opioid receptors kappa, delta and mu
Wash Buffer Ultimate
concentration Tris-HCl 50 mM pH adjusted to 7.4
storage at 4°C

Techniques

1) Preparation of membranes and radioligands

Target Membrane Concentration (µg/well) Radioligand Radioligand concentration (nM) DOR 6.7 [ ^3H ]-DADLE 0.5 MOR 20 [ ^3H ]DAMGO 0.5 Kor 6.7 [ ³ H] Diprenophrine 0.3

2) Preparation of connections

Target Initial conc. compounds in the plate with the original dilution
(mm) Final initial conc. in assay plate (nM) Control: initial conc. in the tablet with the original dilution
(mm) Control: final initial conc. in tablet for analysis
(nM) Concentration of NSB Compound in Assay Plate DOR 2 10000 0.02 100 naltrindol
(1 µM) MOR 2 10000 0.2 1000 naltrindol
(1 µM) Kor 2 10000 0.2 1000 trans-U-50488
(5 µM)

3) Method of quantitative analysis

1) Dispense 1 µl of the compound of a certain concentration into the assay plate according to the plate chart for non-specific binding. Dispense 1 µl of DMSO (dimethyl sulfoxide) into the assay plate according to the plate chart for total binding.

2) Following the plate map, dispense 99 µl of the original membrane preparation into the plate.

3) Add 100 µl of radioligand.

4) Seal the plates tightly. Incubate for 1 hour at room temperature.

5) Moisten Unifilter-96 GF/C filter plates with 50 µl of 0.3% PEI per well, run for at least 0.5 h at room temperature.

6) After the binding assay is complete, filter the reaction mixture through GF/C plates using a Perkin Elmer Filtermate Harvester, and then wash each plate 4 times with cold wash buffer.

7) Dry the filter plates for 1 hour at 50°C.

8) After drying, seal the bottom of the wells of the filter plates using Perkin Elmer Unifilter-96 sealing pad. Add 50 µl Perkin Elmer Microscint 20 c°C ktail.

Seal the lid of the filter plates with Perkin Elmer TopSeal-A sealing film.

9) On the second day, count the radioactivity of the ³ H retained on the filter using a Perkin Elmer MicroBeta2 plate reader.

10) Perform data analysis using GraphPad Prism 5. Calculate "Inhibition [% of Control]" using the equation: % Inhibition = (1-background analyte subtract/background HC subtract)*100.

results

Control Kor U-50488 DOR naltrindol MOR DAMGO

Table 5a Compound DOR IC50
(nM) Maximum
dose (nM) %Inhibition@
Maximum dose thirty 1854.0 10000 84.3 32 >10000 10000 43.2 25 9540.0 10000 62.0 6 499.5 10000 70.9 23 106.7 10000 95.6 24 2121 10000 48.0 34 >10000 10000 22.7 44 190.1 10000 79.1 45 68.4 10000 73.8 46 1360 10000 72.3 nalmefene 18.1 1000 94.5 naltrindol 0.2 100 98.8

Table 5b Compound Kor IC50
(nM) Maximum
dose (nM) %Inhibition@
Maximum dose thirty 88.7 10000 97.6 32 2116.0 10000 80.1 25 889.3 10000 76.9 6 51.1 10000 99.6 23 11.7 10000 101.6 24 37.5 10000 82.9 34 1767 10000 68.1 44 18.4 10000 99.2 45 5.5 10000 100.1 46 33.85 10000 101.8 nalmefene 2.0 1000 103.0 U-50488 10.1 1000 101.6

Table 5c Compound MOR IC50
(nM) Maximum
dose (nM) %Inhibition@
Maximum dose thirty 9.0 10000 82.5 32 17.4 10000 75.9 25 5.0 10000 87.3 6 14.9 10000 102.5 23 2.8 10000 98.7 34 12.55 10000 92.7 44 3.029 10000 83.1 45 2 10000 93.7 46 4.523 10000 102.3 24 22.4 10000 88.7 nalmefene 0.4 1000 103.5 DAMGO 1.4 1000 99.3

Example 3 Solubility Determination

[00287] A known amount of the test substance (~40 mg) was weighed into a tube, 100 μl of oil was added, and heated to 60°C, and then the system was suspended to achieve equilibrium. Oil was added until the solution became clear or the solubility remained <50 mg/mL. The resulting clear solution was then kept at room temperature (25° C.) for 24 hours to see if a solid precipitate had formed. More oil was added to the tube if the compound precipitated, and then the system was equilibrated again at 1000 rpm at room temperature (25°C). The final concentration was determined using the HPLC method, such as described below in Tables 6a and 6b.

Table 6a Reagent Name Degree of purity Manufacturer Batch number Purified water for HPLC WuXiAppTec N/A* cottonseed oil N/A SIGMA 038K000G cottonseed oil extra pure CRODA 1070292 Sesame oil for HPLC CRODA 1115393 ethanol for HPLC J.T.Baker 155943 AN (acetonitrile) for HPLC Merck I0904530 735

*N/A - no data.

Table 6b Equipment Name Model Manufacturer Serial number Water purification device Milli-Q Direct 8 MILLIPORE PDS-PF-WPE-01 Scales Mettler-Toledo XPR10 Mettler Toledo PDS-PF-BAL-08 Mixer C-MAG MS 10 IKA PDS-PF-ST-01 Scales Mettler-Toledo MX5 Mettler Toledo PDS-PF-BAL-03 HPLC Shimadzu 20AB Shimadzu PDS-PF-HPLC-12 Thermomixer YQH-0623 Eppendorf PDS-PF-TM-02

The HPLC procedure for the analysis of Compounds 6, 12-20 and 36-43 is shown in Table 7.

Table 7 HPLC Method 1 Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10 cm * 4.6 mm, 2.7 µm Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.0 ml/min Injector volume 10 µl Analysis duration 20 minutes Temperature
columns 40°C Wavelength 280 nm Specified Gradient Time
(min) A% B% 0 80 20 9 10 90 9.01 10 90 14 80 20 20 80 20 20.01 Stop

HPLC methods for the analysis of Compounds 10, 21-23, 53, 55, 56, nalmefene and naltrexone are shown in Table 8.

Table 8 HPLC Method 2 Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10cm*4.6mm, 2.7µm Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.0 ml/min Injector volume 10 µl Analysis duration 13 min Temperature
columns 40°C Wavelength 284 nm Specified Gradient Time
(min) A% B% 0 95 5 9 10 90 eleven 10 90 11.01 95 5 13 95 5 13.01 Stop

The HPLC procedure for the analysis of Compounds 3-5, 8, 24-25, 26-34, 44-51, 54, 55, 57, 59 and 60 is shown in Table 9.

Table 9 HPLC Method 3 Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10cm*4.6mm, 2.7µm Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.0 ml/min Injector volume 10 µl Analysis duration 13 min Temperature
columns 40°C Wavelength 280 nm Specified Gradient Time
(min) A% B% 0 90 10 4 35 65 25 20 80 27 10 90 27.01 90 10 thirty 90 10 30.01 Stop

Table 10 Approximate solubility (mg/ml) in excipient Equilibrium solubility (mg/ml) Pace. heating
(°C) Observed temp.
(°C) Compound Castor oil Cottonseed oil Sesame oil ethanol Water 36 >217 >418.7 >411.6 20-30 0.005 40 40 36 N/A >407 >408 N/A N/A 60 25 37 >226 >405.3 >408.4 75-90 0.002 40 40 37 N/A 68.5-82 80-102 N/A N/A 60 25 38 >248 >406.9 230-411 160-200 0.617 40 40 38 N/A N/A N/A N/A 0.21 60 40 39 >218 225-402.2 >400.3 150-200 0.018 40 40 39 N/A 104-139 106-141 N/A N/A 60 25 6 N/A >420.8 >402.8 120-170 0.002 40 40 6 N/A >397 >386 N/A N/A 60 25 41 N/A >407 >403 >201 0.084 40 40 41 N/A <50 <50 N/A N/A 60 25 53 N/A >403.8 >406.3 >404 0.0143 60 40 53 N/A >408.5 >401.9 N/A N/A 60 25 42 N/A <51 <52 78-93 0.245 40 40 42 N/A <26 <25.8 N/A 0.068 60 40 43 N/A <50 <50 >257 0.05 40 40 43 N/A 140-210 133-199 N/A N/A 60 40 43 (in water) N/A N/A N/A N/A 0.048 40 40 eleven N/A >407 >407 >218 0.0005 40 40 eleven N/A <51 <50 N/A N/A 60 25 11 after suspension in heptane N/A N/A N/A N/A 0.0026 N/A N/A 12 N/A 100-120 70-95 65-75 0.043 40 40 12 N/A >398.5 >411.1 N/A N/A 60 40 12 N/A <51 <50 N/A N/A 60 25 12 (in water) N/A N/A N/A N/A 0.026 40 40 54 N/A <51 <50 N/A N/A 60 25 54 (in water) N/A N/A N/A N/A <0.001 40 40 13 N/A >381 >406 >214 0.002 40 40 13 N/A <50 91-121 N/A N/A 60 25 14 N/A <50 <51 69-83 0.01 40 40 14 N/A <25 <25 N/A N/A 60 40 14 (in water) N/A N/A N/A N/A 0.0056 40 40 14 after suspension in heptane N/A N/A N/A N/A <0.0005 N/A N/A 14 after suspension in EtOAc N/A N/A N/A N/A <0.0005 N/A N/A 15 N/A <51 <52 <20 <0.0002 40 40 15 N/A >401 >404 N/A N/A 60 40 15 N/A >402.6 >396.5 N/A N/A 60 25 15 N/A N/A >403.94 N/A N/A 60 25 16 N/A 57-67 68-81 <25 0.2059 40 40 16 N/A 201-403 202-404 N/A 0.011 60 40 17 N/A >416 >416 >209 0.0008 40 40 17 N/A >408 >487 N/A N/A 60 25 18 N/A >403 >403 >209 <0.0002 40 40 18 N/A >400 >402 N/A N/A 60 25 19 N/A 67-80 103-137 27-42 0.0468 40 40 20 N/A >414 >406 157-197 <0.0002 40 40 20 N/A 68-82 80-100 N/A N/A 60 25 61 (Nalme-fen) N/A <25.0 <25.5 102-137 0.025 60 40 62 (Naltrek-son) N/A <26.6 <25.9 103-137 0.295 60 40 56 N/A 25-50 25-50 <40.4 0.0679 60 40 56 N/A <50 <50 N/A N/A 60 25 55 N/A >400.1 >394.8 >401.1 0.0066 60 40 55 N/A <50 <50 N/A N/A 60 25 55 (after slurry in ETOAc) N/A N/A N/A N/A <0.0005 N/A N/A 22 N/A >405.9 >401.9 >397.7 0.0314 60 40 22 N/A 67-80 67-80 N/A N/A 60 25 23 N/A >411.8 >420.7 >419.3 0.0094 60 40 23 N/A >407.7 >423.9 N/A N/A 60 25 10 N/A >420.7 >404.1 >404.7 0.0006 60 40 10 N/A 100-134 <51 N/A N/A 60 25 21 N/A 101-134.8 101-134.5 50-100 0.0006 60 40 50 N/A <51 <52 N/A N/A 60 25 50 (in water) N/A N/A N/A N/A <0.001 40 40 51 N/A <50 <46 N/A N/A 60 25 51 (in water) N/A N/A N/A N/A <0.001 40 40 8* N/A <50 <52 N/A N/A 60 25 8 (in water)* N/A N/A N/A N/A 0.57 40 40 56 NMF palmitate N/A 25-50 25-50 <40.4 0.0066 60 40 56 NMF palmitate N/A <50 <50 N/A N/A 60 25 57 N/A >423.7 >417.3 N/A N/A 60 25 57 (in water) N/A N/A N/A N/A <0.001 40 40 58 N/A 101.8-203.6 99.4-198.8 N/A N/A 60 25 58 (in water) N/A N/A N/A N/A 0.0014 40 40 59 NMF dodeca noate N/A >414 >393.3 N/A N/A 60 25 59 NMF dodeca noate (in water) N/A N/A N/A N/A <0.001 40 40 thirty N/A 252.5-404.06 252.55-398.6 N/A N/A 60 25 30 (in water) N/A N/A N/A N/A <0.001 40 40 31 N/A >416.9 >413.5 N/A N/A 60 25 31 (in water) N/A N/A N/A N/A <0.001 40 40 32 N/A >395.8 >410 N/A N/A 60 25 32 (in water) N/A N/A N/A N/A 0.0328 40 40 33 N/A <50 <51 N/A N/A 60 25 33 (in water) N/A N/A N/A N/A <0.001 40 40 28 N/A <50.2 <50.6 N/A N/A 60 25 28 (in water) N/A N/A N/A N/A <0.001 40 40 24 N/A >403 338-368 N/A N/A 60 25 24 (in water) N/A N/A N/A N/A <0.001 40 40 25 N/A >414 >404 N/A N/A 60 25 25 (in water) N/A N/A N/A N/A <0.001 40 40 34 N/A >404 >405 N/A N/A 60 25 34 (in water) N/A N/A N/A N/A <0.001 40 40 44 N/A >407 208-231 N/A N/A 60 25 44 (in water) N/A N/A N/A N/A <0.001 40 40 45 N/A >402 267-288 N/A N/A 60 25 45 (in water) N/A N/A N/A N/A <0.001 40 40 46 N/A >395.5 202-269 N/A N/A 60 25 46 (in water) N/A N/A N/A N/A <0.001 40 40 29 N/A >410.2 >416.5 N/A N/A 60 25 29 (in water) N/A N/A N/A N/A <0.001 40 40 27 N/A >394.7 >399.2 N/A N/A 60 25 27 (in water) N/A N/A N/A N/A <0.001 40 40 47 N/A >405.5 >404.9 N/A N/A 60 25 47 (in water) N/A N/A N/A N/A <0.001 40 40 48 N/A >410.5 >404.8 N/A N/A 60 25 48 (in water) N/A N/A N/A N/A 0.003 40 40 26 N/A 204.3-239 <354 N/A N/A 60 25 26 (in water) N/A N/A N/A N/A <0.001 40 40 49 N/A <50 <51 N/A N/A 60 25 49 (in water) N/A N/A N/A N/A 0.028 40 40 Solubility determined with a pycnometer Equilibrium solubility
(mg/ml) Pace. heating
(°C) Observed temp.
(°C) 8* N/A N/A >399.8 N/A N/A 60 25 8 (in water)** N/A N/A N/A N/A <0.001 40 40 3 N/A >399.12 >400.7 N/A N/A 60 25 3 (in water) N/A N/A N/A N/A <0.001 40 40 4 N/A >395.3 >400.4 N/A N/A 60 25 4 (in water) N/A N/A N/A N/A <0.001 40 40 5 N/A >401.8 >400.6 N/A N/A 60 25 5 (in water) N/A N/A N/A N/A <0.001 40 40

*Data for Compound 8 from the initial failed run from a non-optimized synthesis;

** Data for compound 8 from the second series obtained after a successfully optimized synthesis.

Example 4: Determination of the stability of the finished dosage form

[00288] Compounds were resuspended in oily vehicles, stored at room temperature for the indicated time period, and analyzed by HPLC. The results are presented as a percentage of the absolute loss, normalized to 30 days. Data have previously been published for nalmefene dodecanoate (Gaekens et al., Journal of Controlled Release 232 (2016) 196-202).

Table 11 Reagents Name Degree of purity Manufacturer Batch number Water for HPLC WuXiAppTec N/A AN for HPLC Merck I0911030735 cottonseed oil N/A SIGMA 038K000G cottonseed oil extra pure CRODA 1070292 Sesame oil N/A ACROS A0377665 Sesame oil for HPLC CRODA 1115393 TFU for HPLC J&K LN20M33 THF
(tetrahydrofuran) for HPLC MACRON 1613729801

Table 12 Equipment Name Model Manufacturer Serial number Water purification device Milli-Q Direct 8 MILLIPORE PDS-PF-WPE-01 Scales Mettler-Toledo XP6 Mettler Toledo PDS-PF-BAL-05 HPLC Shimadzu 20AB Shimadzu PDS-PF-HPLC-12

Table 13 HPLC method Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10cm*4.6mm, 2.7µm Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.2 ml/min Injector volume 10 µl Analysis duration 30 min Temperature
columns 40°C Wavelength 280 nm Diluent THF Specified Gradient Time
(min) A% B% 0 90 10 4 35 65 25 20 80 27 10 90 27.01 90 10 thirty 90 10 30.01 Stop

Table 14 Compound %, Average degradation in cottonseed oil (normalized for 30 days) %, Average degradation in Sesame oil (normalized for 30 days) 29 0.13 0.32 27 0.13 0.32 32 - 0.13 24 0.43 (0.01) 25 0.28 (0.11) 31 1.15 0.08 6 1.48 0.31 15 0.86 0.10 17 1.05 0.26 18 1.25 0.71 23 0.35 0.09 5 0.40 0.28 59 4.60 0.51 34 (0.11) 0.02 44 0.36 (0.08) 45 0.31 0.09 46 0.04 Not tested 47 0.43 Not tested 48 0.60 Not tested 3 0.04 0.07

Example 4: Determination of drug substance stability

[00289] Compounds were stored at room temperature for the indicated period of time and analyzed using HPLC. The results are presented as a percentage of the absolute loss, normalized to 30 days. Data have previously been published for nalmefene dodecanoate (Gaekens et al., Journal of Controlled Release 232 (2016) 196-202).

Table 15 Reagent Name Degree of purity Manufacturer Batch number Purified water for HPLC WuXiAppTec N/A AN for HPLC Merck JA056730 TFU for HPLC J&K LN20M33 THF for HPLC MACRON 1613729801

Table 16 Equipment Name Model Manufacturer Serial number Water purification device Milli-Q Direct 8 MILLIPORE PDS-PF-WPE-01 Scales Mettler-Toledo XPR10 Mettler Toledo PDS-PF-BAL-08 Scales Mettler-Toledo MX5 Mettler Toledo PDS-PF-BAL-03 HPLC Shimadzu 20AB Shimadzu PDS-PF-HPLC-12

Table 17 HPLC procedure for the analysis of compounds 10, 11, 21, 36 and 53 HPLC Method 1 Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10cm*4.6mm, 2.7um Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.2 ml/min Injector volume 10 µl Analysis duration 32 min Column temperature 40°C Diluent AN:H ₂ O (3:1) Wavelength 280 nm Specified Gradient Time
(min) A% B% 0 90 10 4 40 60 25 5 95 27 5 95 27.01 90 10 32 90 10 32.01 Stop

Table 18 HPLC procedure for the analysis of compounds 14, 19 and 55 HPLC Method 2 Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10 cm * 4.6 mm, 2.7 µm Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.2 ml/min Injector volume 10 µl Analysis duration 32 min Column temperature 40°C Diluent AN:H ₂ O (3:1) Wavelength 280 nm Specified Gradient Time
(min) A% B% 0 90 10 4 35 65 25 20 80 27 10 90 27.01 90 10 thirty 90 10 thirty Stop

Table 19. HPLC procedure for the analysis of compounds 1, 3-9, 15, 17, 18, 23-27, 29, 31, 32, 34, 35, 44-48, 57 and 59 HPLC Method 3 Apparatus Shimadzu 20AB HPLC Column Ascentis Express C18, 10 cm *4.6 mm, 2.7 µm Gradient A = 0.1% TFA/H ₂ O, B = 100% AN Flow rate 1.2 ml/min Injector volume 10 µl Analysis duration 32 min Column temperature 40°C Diluent THF Wavelength 280 nm Specified Gradient Time
(min) A% B% 0 90 10 4 35 65 25 20 80 27 10 90 27.01 90 10 thirty 90 10 thirty Stop

Table 20 Compound Purity (initial) Purity (after storage at room temperature) % degradation % degradation,
normalized for 30 days 36 97.70% 81.19% 16.51% 5.27% 6 99.00% 99.56% -0.56% -0.18% 53 97.00% 89.90% 7.10% 3.95% eleven 98.20% 98.96% -0.76% -0.28% 14 97.01% 97.90% -0.89% -0.33% 15 98.40% 98.64% -0.24% -0.10% 17 96.30% 98.38% -2.08% -0.84% 18 98.44% 98.43% 0.01% 0.00% 19 99.70% 100.00% -0.30% -0.13% 55 98.80% 98.89% -0.09% -0.04% 23 97.30% 99.97% -2.67% -1.31% 10 97.40% 80.74% 16.66% 8.19% 21 97.60% 98.81% -1.21% -0.60% 57 99.60% 99.84% -0.24% -0.17% 59 97.41% 99.09% -1.68% -1.23% 31 98.80% 98.80% 0.00% 0.00% 32 99.00% 99.22% -0.22% -0.23% 24 99.00% 98.86% 0.14% 0.11% 25 99.00% 98.33% 0.67% 0.38% 34 98.00% 97.87% 0.13% 0.07% 44 98.00% 98.40% -0.40% -0.22% 45 98.00% 98.49% -0.49% -0.28% 46 97.00% 96.97% 0.03% 0.02% 29 99.00% 99.43% -0.43% -0.46% 27 98.90% 98.11% 0.79% 0.85% 47 99.00% 99.24% -0.24% -0.29% 48 99.00% 98.89% 0.11% 0.15% 26 99.00% 95.89% 3.11% 0.72% 7 99.00% 99.32% -0.32% -0.10% 8 98.30% 98.48% -0.18% -0.05% 9 99.00% 97.59% 1.41% 0.35% 1 99.00% 99.18% -0.18% -0.07% 3 99.00% 99.62% -0.62% -0.30% 4 99.00% 99.73% -0.73% -0.24% 5 100.00% 100.00% 0.00% 0.00% 35 100.00% 100.00% 0.00% 0.00%

Example 5 Physical Characteristics of the Solid State Drug Substance

генератор - напряжение 40 кВ, ток 40 мА; диапазон сканирования - от 3° до 40°; скорость вращения образца - 15 об/мин; частота сканирования - 10 град./мин. DSC: образец массой ~1 мг исследовали с использованием завальцованного алюминиевого тигля, закрытого крышкой с отверстием, нагреваемого от комнатной температуры до 300°C со скоростью 10°C/мин. TGA: образец массой 2-5 мг помещали в открытый платиновый тигель и нагревали от комнатной температуры до 300°C со скоростью 10°C/мин. Ранее были опубликованы данные для налмефена пальмитата (Gaekens et al., Journal of Controlled Release 232 (2016) 196-202).[00290] Analysis of the physical characteristics of drugs in the solid state was performed using polarized light microscopy (PLM), X-ray powder diffractometry (XRPD), differential scanning calorimetry (DSC) and thermal gravimetric analysis (TGA). PLM: Samples were dispersed in immersion oil and observed using ocular lenses (10x) and objective lenses (20x) under crossed polarizers. XRPD: samples were examined on a diffractometer using the following technique: tube - Cu: K-alpha

generator - voltage 40 kV, current 40 mA; scanning range - from 3° to 40°; sample rotation speed - 15 rpm; scanning frequency - 10 deg./min. DSC: a sample weighing ~1 mg was examined using a rolled aluminum crucible, closed with a lid with a hole, heated from room temperature to 300°C at a rate of 10°C/min. TGA: A 2-5 mg sample was placed in an open platinum crucible and heated from room temperature to 300°C at a rate of 10°C/min. Data have previously been published for nalmefene palmitate (Gaekens et al., Journal of Controlled Release 232 (2016) 196-202).

Table 21. XRPD, TGA and DSC data Compound X-ray powder diffractometry Thermal gravimetric analysis Differential Scanning Calorimetry eleven Crystalline form with clear birefringence Mass loss: 0.30% at 120°C after decomposition One endothermic peak at 56.71°C, which could be the melting point. 12 Crystalline form with clear birefringence Mass loss: 0.19% at 120°C after decomposition One endothermic peak at 51.94°C, which could be the melting point. 14 Weak crystallinity of the compound Mass loss: 1.649% at 120°C after decomposition One endothermic peak at 124.66°C, which could be the melting point. 16 Crystalline form with clear birefringence Mass loss: 0.403% at 120°C after decomposition One endothermic peak at 59.04°C, which could be the melting point. 19 Crystalline form with clear birefringence Mass loss: 0.1623% at 120°C after decomposition One endothermic peak at 56.31°C, which could be the melting point. 56 Crystalline form with clear birefringence Mass loss: 0.1815% at 120°C after decomposition Two endothermic peaks at 48.95°C and 59.75°C. 55 Crystalline form with clear birefringence Mass loss: 0.2597% at 120°C after decomposition One endothermic peak at 48.24°C, which could be the melting point. 10 Crystalline form with clear birefringence Mass loss: 0.3047% at 120°C after decomposition One endothermic peak at 47.18°C, which could be the melting point. 40 Crystalline form with partial birefringence Mass loss: 0.57% at 120°C after decomposition Two endothermic peaks at 107.6°C and 195.0°C. 21 Crystalline form with partial birefringence. Mass loss: 1.133% at 120°C after decomposition One endothermic peak at 70.84°C, which could be the melting point. 31 Crystalline form with clear birefringence Mass loss: 0.7244% at 120°C after decomposition One endothermic peak at 45.15°C, which could be the melting point.

Example 5 Screening for solid state drug polymorphisms

[00291] To identify stable polymorphic forms of drugs in the solid state, approximately 50 mg of the compound (in equivalents of nalmefene or naltrexone) were weighed into test tubes. Then 500 µl of the indicated solvents were added and this suspension was stirred at 700 rpm and 50° C. for 72 hours. Upon receipt of samples in the form of a suspension, the solid was separated by centrifugation (10 minutes, 14000 rpm) and dried in a vacuum oven at 30°C overnight. When the samples were prepared as a solution, a solid was formed by evaporating the solvent (removing the stir bar and covering with perforated aluminum foil, then drying in a vacuum oven at 30° C. overnight). The dry solid was analyzed using XRPD, TGA and DSC. The results are shown in Table 23.

Table 23 Compound Solvents Target conc. visual observation Drying technique and appearance of the substance XRPD data (after drying) Comments (mg/ml) k.t. 50°C eleven Heptane 100 Clear solution Clear solution Evaporation/White Powder Form B The original crystalline form (Form A) appears to be the stable form. Form A was obtained from a solution in MTBE, IPA, EtOAc and acetone by evaporation, the compound obtained from a solution in heptane by evaporation had a different form. MTBE Clear solution Clear solution Evaporation/White Powder Form A Isopropanol Suspension Clear solution Evaporation/White Powder Form A EtOAc Suspension Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder Form A 14 Heptane 100 Suspension Suspension Centrifugation/White Powder Form B The original crystalline form (Form A) appears to be unstable; The same crystalline form obtained from a solution in MTBE, IPA, EtOAc and acetone by evaporation may be unstable. The form obtained from the heptane solution by evaporation is a different form and appears to be more stable MTBE Clear solution Clear solution Evaporation/White Powder Form A Isopropanol Clear solution Clear solution Evaporation/White Powder Form A EtOAc Suspension Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder Form A 16 Heptane 100 Suspension Suspension Centrifugation/White Powder Form A The original crystalline form (Form A) appears to be the stable form. The same crystalline form (form A) was obtained from a solution in MTBE, EtOAc and acetone by evaporation. The same crystalline form (form A) was obtained by suspension in heptane and IPA. MTBE Clear solution Clear solution Evaporation/White Powder Form A Isopropanol Suspension Suspension Centrifugation/White Powder Form A EtOAc Suspension Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder Form A 19 Heptane 100 Suspension Clear solution Evaporation/White Powder Form A The original crystalline form (Form A) appears to be the stable form. The same crystalline form (form A) was obtained from a solution in heptane, MTBE, IPA, EtOAc and acetone by evaporation. MTBE Clear solution Clear solution Evaporation/White Powder Form A Isopropanol Suspension Clear solution Evaporation/White Powder Form A EtOAc Suspension Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder Form A 55 Heptane 100 Suspension Clear solution Evaporation/White Powder Form B It cannot be determined which crystal form is more stable. The same crystalline form (form B) was obtained from a solution in heptane, MTBE, IPA, EtOAc and acetone by evaporation. MTBE Clear solution Clear solution Evaporation/White Powder Form B Isopropanol Suspension Clear solution Evaporation/White Powder Form B EtOAc Suspension Clear solution Evaporation/White Powder Form B Acetone Suspension Clear solution Evaporation/White Powder Form B 10 Heptane 100 Suspension Suspension Centrifugation/White wax Form A The original crystalline form (Form A) is the stable form. The same crystalline form (form A) was obtained from a solution in MTBE, IPA, EtOAc and acetone by evaporation. Form A was also prepared by suspension in heptane. MTBE Clear solution Clear solution Evaporation/White wax Form A Isopropanol Suspension Clear solution Evaporation/White wax Form A EtOAc Suspension Clear solution Evaporation/White wax Form A Acetone Suspension Clear solution Evaporation/White wax Form A 21 Heptane 100 Suspension Clear solution Evaporation/White Powder Form A The original crystalline form (Form A) is the stable form. The same crystalline form (form A) was obtained from a solution in heptane, MTBE, IPA, EtOAc and acetone by evaporation. MTBE Clear solution Clear solution Evaporation/White Powder Form A Isopropanol Suspension Clear solution Evaporation/White Powder Form A EtOAc Suspension Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder Form A 31 Heptane 100 Suspension Clear solution Evaporation/White Powder Form A The original crystalline form (Form A) is likely to be the stable form. The same crystalline form (form A) was obtained from a solution in MTBE, IPA, EtOAc and acetone by evaporating MTBE Suspension Clear solution Evaporation/White Powder Form A Isopropanol Clear solution Clear solution Evaporation/White Powder Form A EtOAc Clear solution Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder Form A 40 Heptane 100 Suspension Suspension Centrifugation/White Powder Form A The original crystalline form (Form A) is the stable form. The same crystalline form (form A) was obtained by suspension in heptane and from a solution in EtOAc by evaporation. The compound obtained from a solution in MTBE, IPA and acetone by evaporation had an amorphous form. MTBE Clear solution Clear solution Evaporation/White Powder amorphous form Isopropanol Suspension Clear solution Evaporation/Oil amorphous form EtOAc Suspension Clear solution Evaporation/White Powder Form A Acetone Suspension Clear solution Evaporation/White Powder amorphous form

III. Obtaining pharmaceutical dosage forms

Example 1: Capsule for oral administration

[00292] The compound of Table 1, or a pharmaceutically acceptable salt thereof, was used as the active ingredient. A capsule for oral administration is prepared by mixing 1-1000 mg of the active ingredient with starch or other suitable powder mixture. The resulting mixture is formulated into an oral dosage unit form, such as a hard gelatin capsule, which is suitable for oral administration.

Example 2: Solution for injection

[00293] The compound from Table 1, or a pharmaceutically acceptable salt thereof, was used as the active ingredient, the preparation was obtained as a solution in sesame oil, cottonseed oil, castor oil or other pharmaceutically acceptable lipophilic excipient containing the active ingredient preferably at a concentration of more than 100 mg / ml. The resulting solution was administered by intramuscular injection.

Compounds were resuspended in a volume of 1 ml at the indicated concentrations (in mg/ml base equivalents) by stirring on a magnetic stirrer (1000 rpm) at 60°C until a homogeneous transparent solution was obtained, then cooled to room temperature and stored in the dark place. The appearance of the oil preparations was evaluated and recorded at room temperature (25°C) at the beginning of the observation, after 2 hours and 24 hours. Samples for "Compound Quantification" were taken immediately after resuspension, 2 hours and 24 hours and analyzed by HPLC, the actual concentration was determined based on the calibration curve (Compound Quantification = Concentration (measured by HPLC)/Actual concentration (by mass) x 100%). Purity was calculated at the indicated time points as the percentage of the area under the curve of the main peak on the HPLC spectrogram. The suitability of preparations for administration using a syringe was assessed by pumping them through a 21 gauge needle. For some of these samples, after storage for 7 months at 40°C and 75% relative humidity, the appearance was evaluated, quantitative analysis and purity analysis were performed. The results are shown in Table 24.

Table 24 Appearance Compound Quantification Purity Suitability of drugs for administration using a syringe (21-gauge needle) Follow-up at room temperature Compound Carrier Volume Target conc. (mg/ml) in equiv. foundation Outgoing In 2 hours after 24 hours Initial, % In 2 hours, % after 24 hours, % Initial, % In 2 hours, % after 24 hours, % Outcome In 2 hours after 24 hours 6 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 102.26 102.32 99.30 99.48 99.46 99.37 suitable suitable suitable Remains in solution Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 99.82 99.66 102.90 99.37 99.35 99.44 suitable suitable suitable Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 98.06 97.94 98.43 99.37 99.39 99.37 suitable suitable suitable 15 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 107.68 109.70 109.70 96.38 96.44 96.34 suitable suitable suitable Remains in solution Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 101.55 108.28 103.00 96.44 96.38 96.45 suitable suitable suitable Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 93.52 99.92 94.15 96.42 96.28 96.41 suitable suitable suitable 17 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 98.80 98.66 98.29 98.61 98.64 98.61 suitable suitable suitable Remains in solution Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 105.43 105.79 105.65 98.61 98.60 98.63 suitable suitable suitable Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 98.51 98.82 97.58 98.63 98.60 98.59 suitable suitable suitable 18 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 97.56 97.79 97.85 99.15 99.05 99.07 suitable suitable suitable Remains in solution Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 99.00 100.43 99.52 98.93 99.06 98.84 suitable suitable suitable Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 98.54 98.70 98.77 99.17 99.13 99.14 suitable suitable suitable 23 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 97.22 97.54 97.10 99.50 99.48 99.47 suitable suitable suitable Remains in solution Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 100.34 100.25 99.51 99.49 99.48 99.50 suitable suitable suitable Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 97.54 97.92 95.81 99.51 99.48 99.49 suitable suitable suitable 57 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 104.75 104.82 104.02 99.55 99.49 99.54 suitable suitable suitable Insoluble matter: large sediment/phase separation after 23 days Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 102.67 102.47 100.66 99.69 99.67 99.65 suitable suitable suitable Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 101.03 101.18 100.34 99.63 99.59 99.63 suitable suitable suitable 31 Sesame oil + 10 µl benzyl alcohol 1 ml 200 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 101.90 100.35 N/A 99.29 99.28 N/A suitable suitable suitable Insoluble matter: large sediment/phase separation after 24 hours Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil white wax 105.84 95.68 N/A 99.14 99.31 N/A suitable suitable N/A Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil white wax 99.16 95.81 N/A 99.23 99.25 N/A suitable suitable N/A 32 Sesame oil + 10 µl benzyl alcohol 1 ml 300 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 95.61 96.59 95.42 98.78 98.63 98.78 suitable suitable suitable Insoluble matter: large sediment/phase separation after 24 hours Sesame oil + 10 µl benzyl alcohol 1 ml 350 Low viscosity orange transparent oil Low viscosity orange transparent oil white wax 102.17 103.30 N/A 98.79 98.76 N/A suitable suitable N/A Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil white wax 97.21 97.10 N/A 98.80 98.78 N/A suitable suitable N/A 24 Cottonseed oil + 10 µl Benzyl alcohol 1 ml 400 Low viscosity clear yellow oil Low viscosity clear yellow oil Low viscosity clear yellow oil 107.62 99.58 100.58 99.02 98.86 98.93 suitable suitable suitable Remains in solution 29 Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 103.07 104.83 103.35 99.21 99.21 99.28 suitable suitable suitable Drug substance is unstable in oily solution when stored for 4 months at 40°C/75% RH 27 Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 95.52 98.43 96.46 99.33 99.30 99.09 suitable suitable suitable Insoluble matter: large sediment/phase separation after 90 days 47 Sesame oil + 10 µl benzyl alcohol 1 ml 400 Low viscosity orange transparent oil Low viscosity orange transparent oil Low viscosity orange transparent oil 102.41 101.60 101.72 99.37 99.31 99.26 suitable suitable suitable Insoluble matter: large sediment/phase separation after 24 hours

Table 25A Compound Carrier Sample volume Target conc. (mg/ml) amount of nalmefene Appearance Compound Quantification original In 2 hours after 24 hours after 7 months storage at 40°C/75% RH Outgoing In 2 hours after 24 hours after 7 months storage at 40°C/75% RH -1 after 7 months storage at 40°C/75% RH -2 after 7 months storage at 40°C/75% RH -3 25 Sesame oil + 20 µl Benzyl alcohol 1+1 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil Low-viscosity cloudy oil, immiscible fractions possible 102.48% 102.90% 108.25% 98.02% 94.38% 98.31% 34 Sesame oil + 20 µl Benzyl alcohol 1+1 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil Yellow cloudy oil, possible immiscible drops of oil 101.77% 102.07% 101.11% 102.01% 99.07% 97.99% 44 J&K Cottonseed Oil + 10µl Benzyl Alcohol 1 ml 350 Low viscosity clear yellow oil Low viscosity clear yellow oil Low viscosity clear yellow oil 101.41% 102.44% 102.32% 45 J&K Cottonseed Oil + 20µl Benzyl Alcohol 1+1 ml 300 Light yellow clear oil Light yellow clear oil Light yellow clear oil Yellow cloudy oil, small drops of oil possible 98.86% 98.68% 98.64% 100.57% 99.54% 98.78% 46 J&K Cottonseed Oil + 20µl Benzyl Alcohol 1+1 ml 300 Light yellow cloudy oil Light yellow cloudy oil Light yellow cloudy oil Yellow cloudy inhomogeneous oil with pronounced immiscible oil drops 98.94% 99.28% 99.00% 106.24% 108.43% 107.85% 48 Sesame oil + 10 µl Benzyl alcohol 1 ml 400 Light-viscosity cloudy oil Light-viscosity cloudy oil Light-viscosity cloudy oil 97.50% 96.63% 96.39%

Table 25B Compound Purity Suitability of drugs for administration using a syringe (21-gauge needle) Follow-up at room temperature Follow-up after 7 months storage at 40°C/75% RH Initial In 2 hours after 24 hours after 7 months storage at 40°C/75% RH -1 after 7 months storage at 40°C/75% RH -2 after 7 months storage at 40°C/75% RH -3 Initial In 2 hours after 24 hours 25 98.36% 98.64% 98.57% 97.07% 97.13% 97.07% fit fit fit Remains in solution No clearly visible particles under PLM microscopy 34 98.59% 98.58% 98.50% 99.35% 99.44% 99.46% fit fit fit Remains in solution No clearly visible particles under PLM microscopy 44 98.68% 98.41% 98.73% fit fit fit Insoluble matter: significant turbidity with noticeable solid sediment after 20 days 45 99.58% 101.17% 100.62% 94.60% 94.68% 94.67% fit fit fit Remains in solution Possible small visible particles under PLM microscopy 46 97.96% 97.96% 97.95% 95.48% 95.42% 95.54% fit fit fit Insoluble matter: significant turbidity with noticeable solid sediment after 20 days Expressed particles in PLM microscopy 48 98.97% 98.80% 98.53% fit fit fit Insoluble Matter: Severe turbidity with noticeable solid sediment after 170 days. Crystalline particles under PLM microscopy. The drug substance turned from a colorless oil to a white wax in 4 weeks.

Table 26A Compound Carrier Sample volume Target conc. (mg/ml) amount of nalmefene Appearance original after 24 hours after 3 months storage at 40°C/75% RH after 6 months storage at 40°C/75% RH 6 Sesame oil + 50 µl Benzyl alcohol 5 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil (at 25°C) Low viscosity orange clear oil 15 Sesame oil + 50 µl Benzyl alcohol 5 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil 17 Sesame oil + 50 µl Benzyl alcohol 5 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil 18 Sesame oil + 50 µl Benzyl alcohol 5 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil 9 Sesame oil + 50 µl Benzyl alcohol 5 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil Low viscosity orange clear oil 59 Sesame oil + 10 µl Benzyl alcohol 5 ml 86 Low viscosity orange clear oil Low viscosity orange clear oil 24 Cottonseed oil + 200 µl Benzyl alcohol 20 ml 400 Pale yellow oil 45 Sesame oil + 1% benzyl alcohol 20 ml 400 clear oil Sesame oil + 1% benzyl alcohol 1 ml 400 clear oil Sesame oil + 1% benzyl alcohol 1 ml 400 clear oil Sesame oil + 1% benzyl alcohol 1 ml 400 clear oil 34 Sesame oil + 100 µl Benzyl alcohol 10 ml 400 brown oil 7 Sesame oil + 1% benzyl alcohol 5+1+1+1 ml 400 Nearly clear solution 43 Sesame oil + 1% benzyl alcohol 10 ml 400 brown oil 9 Sesame oil + 1% benzyl alcohol 5 ml 400 Nearly clear solution 1 ml 1 ml 1 ml 1 Sesame oil + 1% benzyl alcohol 5+1+1 ml 400 Slightly cloudy oil 3 Sesame oil + 1% benzyl alcohol 5+5 ml 400 Low viscosity orange clear oil Low viscosity orange clear oil (after 6 hours) 4 Sesame oil + 1% benzyl alcohol 5+1+1 ml 400 Nearly clear solution 5 Sesame oil + 1% benzyl alcohol 1+1+1 ml 400 Low viscosity orange clear oil 35 Sesame oil + 1% benzyl alcohol 10 ml 400 Nearly clear oil

Table 26B Compound Compound Quantification Initial-1 Initial-2 Initial-3 after 24 hours -1 after 24 hours -2 after 24 hours -3 after 3 months storage at 40°C/75% RH -1 after 3 months storage at 40°C/75% RH -2 after 3 months storage at 40°C/75% RH -3 after 6 months storage at 40°C/75% RH -1 after 6 months storage at 40°C/75% RH -2 after 6 months storage at 40°C/75% RH -3 6 103.19% 106.47% 104.72% 100.81% 100.60% 101.40% 102.15% (25C) 101.9%
(25C) 99.07%
(25C) 101.25% 101.08% 101.24% 15 99.37% 98.69% 101.17% 97.06% 101.32% 99.43% 97.75% 97.98% 97.53% 101.91% 101.82% 102.09% 17 98.84% 99.62% 99.49% 98.02% 98.22% 98.07% 101.42% 101.39% 101.30% 103.37% 103.24% 103.18% 18 93.67% 98.55% 96.11% 95.75% 95.77% 93.03% 102.46% 103.29% 102.94% 99.73% 100.49% 100.23% 9 102.27% 100.89% 101.56% 99.70% 97.28% 90.59% 101.74% 102.03% 101.57% 98.23% 98.80% 93.52% 59 102.52% 102.59% 102.71% 103.05% 102.47% 102.90% 24 112.73% 105.24% 103.76% 45 98.12% 98.99% 100.30% 100.05% 101.02% 99.92% 99.86% 101.14% 99.39% 99.91% 99.03% 99.99% 34 96.78% 97.68% 98.16% 7 99.73% 100.16% 101.37% 43 99.27% 99.39% 97.86% 9 99.23% 99.79% 99.83% 99.35% 98.76% 98.66% 98.93% 100.93% 103.17% 109.00% 109.82% 111.59% 1 110.88% 111.97% 110.59% 3 N/A N/A N/A 97.91% (after 6 days) 97.55% (after 6 days) 94.13% (after 6 days) 4 100.31% 97.12% 97.52% 5 96.64% 95.72% 96.29% 35 98.21% 96.79% 99.37%

Table 26C Compound Purity Suitability of drugs for administration using a syringe (21-gauge needle) Follow-up at room temperature Exodus-1 Exodus-2 Exodus-3 after 24 hours -1 after 24 hours -2 after 24 hours -3 after 3 months storage at 40°C/
75% RHV-1 after 3 months storage at 40°C/
75% RHV-2 after 3 months storage at 40°C/
75% RHV-3 after 6 months storage at 40°C/
75% RHV-1 after 6 months storage at 40°C/
75% RHV-2 after
6 months storage
at 40°C/
75% RHV-3 Outcome after 24 hours in 3 months in 6 months 6 99.48% 99.46% 99.37% 99.36% 99.35% 99.44% 99.29% (25°C) 99.81%
(25°C) 98.75%
(25°C) 98.88% 98.86% 98.89% suitable suitable suitable suitable Remains in solution 15 96.87% 97.19% 96.88% 96.80% 96.39% 96.84% 96.86% 96.57% 96.70% 96.20% 96.18% 96.21% suitable suitable suitable suitable Remains in solution 17 98.49% 98.70% 98.78% 98.45% 98.65% 98.82% 98.52% (25°C) 98.29% (25°C) 98.92% (25°C) 98.74% 98.75% 98.70% suitable suitable suitable suitable Remains in solution 18 99.32% 99.09% 98.97% 99.00% 99.14% 99.07% 98.38% 98.39% 98.39% 98.94% 98.54% 98.71% suitable suitable suitable suitable Remains in solution 9 99.38% 99.36% 99.49% 99.33% 99.56% 99.44% 99.44% 99.17% 99.49% 99.56% 99.53% 99.23% suitable suitable suitable suitable Remains in solution 59 99.00% 98.88% 98.94% 99.03% 98.87% 98.91% suitable suitable Remains in solution 24 99.63% 99.62% 99.64% suitable Remains in solution 45 99.73% 99.71% 99.68% suitable Remains in solution 99.48% 99.38% 99.58% N/A 99.86% 99.63% 99.68% N/A 99.73% 99.57% 99.78% N/A 34 95.93% 95.84% 95.91% suitable The purity of the compound in the finished dosage form was 95.9%. After synthesis, the purity was 99%. Precipitation was observed after 55 days 7 99.34% 99.31% 99.23% suitable Remains in solution 43 98.60% 98.61% 98.49% suitable Remains in solution 9 98.26% 98.14% 97.98% suitable Appearance: cloudy, homogeneous, possible formation of a solution after ~24 hours Visible solid precipitate after ~24 hours Visible solid precipitate after ~24 hours Apparent precipitation from solution after ~24 hours 1 100.00% 100.00% 100.00% suitable Remains in solution 3 N/A N/A N/A 99.5% (after 6 days) 99.54% (after 6 days) 99.34% (after 6 days) suitable Remains in solution 4 99.65% 99.66% 99.76% suitable Remains in solution 5 99.72% 99.71% 99.72% suitable Remains in solution 35 100.00% 100.00% 100.00% suitable Remains in solution

IV. Pharmacokinetic evaluation

Example 1: Pharmacokinetic studies in rats

Target

[00294] The purpose of this study was to determine the pharmacokinetic parameters of the test compounds in plasma after intramuscular administration to male Sprague Dawley rats (n=3, unless otherwise indicated).

Acclimatization/Quarantine

[00295] The general health of the animals was assessed, before being used in the study, the animals were placed for acclimatization for a period of at least 3 days.

Animal keeping

[00296] Animals were kept indoors during acclimatization and kept separately during the study. Animals were kept in a climate controlled room (target conditions: temperature from 18 to 26°C, relative humidity from 30 to 70%, artificial light for 12 hours and darkness for 12 hours). Carried out daily monitoring of temperature and relative humidity. Animals' access to water was not restricted.

Animal body weight and clinical observation

[00297] Body weight was determined prior to selection for use in the study and on the day of dosing. Body weight was monitored every week.

Extensive clinical observation, including observation of behavior and activity, reflection, respiration, skin and coat condition, facial expressions, genitourinary system, and the appearance of significant damage to other systems, was performed on the day of dose and each time samples were taken.

Dose Administration

[00298] A dose of 400 mg base equivalents/ml in sesame oil + 1% benzyl alcohol (unless otherwise indicated) was administered by intramuscular injection. The volume of the dose was administered depending on the body weight of the animal, which was determined on the morning of the day of dosing.

Sampling

[00299] Blood samples were taken from each animal (approximately 0.2 ml at each of the time points) by puncture of the jugular vein, the samples were placed in pre-chilled plastic microcentrifuge tubes containing sodium fluoride/potassium oxalate (5 μl, 160 mg/ml, NaF/KO=1/3), 5% PMSF (100 mM in ethanol) as stabilizer and EDTA-K ₂ (4 μl) as anticoagulant, which were kept on liquid ice until centrifugation.

Getting Plasma

[00300] Each blood sample obtained was centrifuged for 4 minutes at 4°C and 10,000 rpm to collect plasma. Plasma was collected and transferred to a pre-labeled PP tube placed in dry ice at each time point, and immediately precipitated using AN at a ratio of 1:4 (plasma:AN). Centrifuged again (10 minutes, 12,000 rpm) and the supernatant was collected.

After sampling was completed and prior to bioassay, the entire supernatant was stored at approximately -80°C.

Bioanalytical methodology and sample analysis

[00301] LC-MS/MS techniques have been developed to quantify a test compound in a biological environment. This method used a calibration curve with 8 non-zero calibration standards, including LLOQ (0.05 ng/ml). Sample analysis was performed simultaneously with a set of calibration standards and two sets of QC samples using this LC-MS/MS method.

Data analysis

[00302] Plasma concentration versus time data was analyzed by non-compartmental analysis using Phoenix WinNonlin 6.3 software. Received the values of C _max , T _max , T _1/2 , AUC _(0-t) , AUC _(0-inf) , MRT _(0-t) , MRT _(0-inf) and graphs of plasma concentration versus time.

[00303] The dose of nalmefene dodecanoate was determined by allometric scaling per rat of dog doses known from previously published data (Gaekens et al., Journal of Controlled Release 232 (2016) 196-202). The terminal half-life of the active metabolite of the selected compounds was determined and these data were used to estimate the period during which the plasma concentration of the active metabolite is above the minimum effective concentration.

Table 27 Compound Nominal dose
(mg/kg) Carrier Half-life t _1/2 (h)
prodrugs Terminal half-life t _1/2 (h)
nalmefena AUC _0-inf
(ng x h/ml)
prodrugs AUC _0-inf
(ng x h/ml)
nalmefena Nalmefene HCl - IR 0.80 saline - 0.87 - 66.0 59 17 Sesame oil+1% benzyl alcohol 15.2 248 10.4 1026 6 80 Sesame oil+1% benzyl alcohol 340 569 78.9 3576 6 123 Sesame oil+1% benzyl alcohol 213 425 620 14704 6 165 Sesame oil+1% benzyl alcohol 204 491 637 18876 15 80 Sesame oil+1% benzyl alcohol 1553 639 679 3046 15 123 Sesame oil+1% benzyl alcohol 884 993 2574 7232 15 165 Sesame oil+1% benzyl alcohol 500 451 2789 14019 17 200 Sesame oil+1% benzyl alcohol No prodrug found 266 No prodrug found 16178 18 80 Sesame oil+1% benzyl alcohol No prodrug found 599 No prodrug found 8217 18 123 Sesame oil+1% benzyl alcohol No prodrug found 3409 No prodrug found 13658 18 200 Sesame oil+1% benzyl alcohol No prodrug found 847 No prodrug found 15104 23
(metabolite
naltrexone) 200 Sesame oil+1% benzyl alcohol No prodrug found Not rated No prodrug found 27105
(metabolite
naltrexone) 24 80 Cottonseed oil + 1% benzyl alcohol 398 1093 147 5584 24 123 Cottonseed oil + 1% benzyl alcohol 717 445 547 11283 24 165 Cottonseed oil+1% benzyl alcohol 851 458 453 19031 29 165 Sesame oil+1% benzyl alcohol 175 911 74.8 17313
(metabolite
naltrexone)

Throughout the study, none of the rats had a negative effect on body weight or adverse clinical observations.

[00304] Nalmefene time vs. concentration data for nalmefene HCL (1 mg/mL) at 80 mg/kg are shown in Table 28a.

Table 28a Time (h) Average concentration
nalmefene (ng/ml) 1.00 29.4 2.00 2.09 4.00 0.212 8.00 0.0178 12 ND* 24.0 ND

*Not registered.

[00305] Nalmefene time vs. concentration data for a 17 mg/kg dose of Compound 59 (nalmefene dodecanoate, at 86 mg/mL) are shown in Table 28b.

Table 28b Time (h) Average concentration
nalmefene (ng/ml) 1.00 7.85 2.00 8.10 4.00 7.45 8.00 6.85 24.0 6.96 48.0 7.92 72.0 5.77 144 1.59 312 0.403 480 0.235 648 0.145

[00306] Nalmefene time vs. concentration data for the 80 mg/kg dose of Compound 6 are shown in Table 29.

Table 29 Time (h) Average concentration
nalmefene (ng/ml) 1.00 9.51 2.00 7.11 4.00 5.48 8.00 5.09 24.0 5.74 48.0 8.48 72.0 7.18 144 3.73 312 3.08 480 2.49 648 2.09 816 1.80 984 1.39 1152 1.14 1320 0.833 1488 0.413

[00307] Nalmefene time vs. concentration data for a 123 mg/kg dose of Compound 6 are shown in Table 30.

Table 30 Time (h) Average concentration
nalmefene (ng/ml) 0.250 43.8 1.00 36.3 2.00 24.0 4.00 16.7 8.00 12.1 24.0 11.0 48.0 14.2 72.0 10.7 144 9.17 312 13.6 480 10.6 648 11.1 816 8.8 984 6.5 1152 4.0 1320 3.2 1488 1.6 1656 1.7 1824 1.2 1992 0.9 2160 0.8 2328 0.8 2496 0.6 2664 0.5

[00308] Nalmefene time vs. concentration data for a 165 mg/kg dose of Compound 6 are shown in Table 31.

Table 31 Time (h) Average concentration
nalmefene (ng/ml) 0.250 50.4 1.00 48.0 2.00 29.2 4.00 19.3 8.00 13.9 24.0 12.3 48.0 14.4 72.0 12.4 144 10.7 312 14.5 480 18.1 648 17.1 816 14.2 984 10.5 1152 6.2 1320 4.9 1488 2.8 1656 2.3 1824 1.8 1992 1.3 2160 1.3 2328 1.1 2496 0.9 2664 0.7

[00309] Nalmefene time vs. concentration data for the 80 mg/kg dose of Compound 15 are shown in Table 32.

Table 32 Time (h) Average concentration
nalmefene (ng/ml) 1.00 7.07 2.00 4.21 4.00 2.10 8.00 1.42 24.0 1.98 48.0 2.39 72.0 3.25 144 2.32 312 2.18 480 3.03 648 2.15 816 0.944 984 0.714 1152 0.745 1320 0.663 1488 0.706 1656 0.697 1824 0.514 1992 0.322 2160 0.441

[00310] Nalmefene time vs. concentration data for the 123 mg/kg dose of Compound 15 are shown in Table 33.

Table 33 Time (h) Average concentration
nalmefene (ng/ml) 0.250 18.7 1.00 20.4 2.00 13.4 4.00 6.20 8.00 2.74 24.0 1.54 48.0 1.89 72.0 2.00 144 2.97 312 5.64 480 7.88 648 6.60 816 5.33 984 3.87 1152 2.41 1320 2.10 1488 2.33 1656 2.01 1824 1.93 1992 1.50 2160 1.25 2328 1.01 2496 1.18 2664 0.831

[00311] Nalmefene time vs. concentration data for a 165 mg/kg dose of Compound 15 are shown in Table 34.

Table 34 Time (h) Average concentration
nalmefene (ng/ml) 0.250 30.3 1.00 24.1 2.00 15.3 4.00 7.60 8.00 2.92 24.0 1.65 48.0 2.43 72.0 2.60 144 3.11 312 6.45 480 13.2 648 12.5 816 7.92 984 6.30 1152 5.30 1320 4.54 1488 3.62 1656 3.24 1824 3.38 1992 2.16 2160 1.62 2328 0.966 2496 0.831 2664 0.903

[00312] Nalmefene time vs. concentration data for the 200 mg/kg dose of Compound 17 are shown in Table 35.

Table 35 IM Time (h) Average concentration
nalmefene (ng/ml) 1.00 249 2.00 299 4.00 364 8.00 340 24.0 157 48.0 80.0 72.0 44.3 144 15.5 312 5.09 480 3.04 648 3.37 816 1.90 984 1.29 1152 0.737 1320 0.375 1488 0.309

[00313] Nalmefene time vs. concentration data for the 80 mg/kg dose of Compound 18 are shown in Table 36.

Table 36 Time (h) Average concentration
nalmefene (ng/ml) 0.250 5.13 1.00 14.9 2.00 22.9 4.00 39.9 8.00 35.6 24.0 36.6 48.0 32.7 72.0 28.2 144 21.5 312 5.60 480 2.99 648 1.58 816 1.29 984 1.34 1152 0.99 1320 0.75 1488 0.505 1656 0.465 1824 0.412 1992 0.383

[00314] Nalmefene time vs. concentration data for the 123 mg/kg dose of Compound 18 are shown in Table 37.

Table 37 Time (h) Average concentration
nalmefene (ng/ml) 0.250 3.98 1.00 15.6 2.00 24.4 4.00 43.5 8.00 43.7 24.0 39.2 48.0 29.8 72.0 26.5 144 15.9 312 4.21 480 5.59 648 4.73 816 4.01 984 4.26 1152 3.14 1320 3.10 1488 2.44 1656 2.55 1824 2.55 1992 1.38 2160 2.52 2328 3.11 2496 1.92 2664 2.27

[00315] Nalmefene time vs. concentration data for a 200 mg/kg dose of Compound 18 are shown in Table 38.

Table 38 Time (h) Average concentration
nalmefene (ng/ml) 1.00 32.3 2.00 59.0 4.00 64.8 8.00 76.8 24.0 54.2 48.0 46.7 72.0 38.4 144 29.0 312 13.4 480 5.17 648 5.51 816 3.54 984 2.90 1152 2.80 1320 2.05 1488 1.77 1656 1.40 1824 1.68 1992 1.18 2160 1.42 2328 1.43 2496 0.781 2664 1.18 2832 2.34

[00316] Naltrexone time vs. concentration data for the 200 mg/kg dose of Compound 23 are shown in Table 39.

Table 39 Time (h) Average concentration
naltrexone (ng/ml) 1.00 32.3 2.00 655 4.00 727 8.00 603 24.0 384 48.0 181 72.0 109

[00317] Nalmefene time vs. concentration data for Compound 24 at 80 mg/kg are shown in Table 40.

Table 40 Time (h) Average concentration
nalmefene (ng/ml) 0.250 1.93 1.00 2.94 2.00 4.06 4.00 4.69 8.00 4.86 24.0 5.47 48.0 5.43 72.0 5.46 144 3.87 312 3.87 480 4.60 648 5.29 816 3.16 984 2.32 1152 2.14 1320 1.45 1488 1.25 1656 1.05 1824 1.14 1992 1.09 2160 0.971 2328 0.798 2496 0.788 2664 0.719

[00318] Nalmefene time vs. concentration data for Compound 24 at 123 mg/kg are shown in Table 41.

Table 41 Time (h) Average concentration
nalmefene (ng/ml) 0.250 1.89 1.00 2.38 2.00 3.28 4.00 4.14 8.00 4.32 24.0 6.20 48.0 5.56 72.0 5.03 144 4.54 312 5.42 480 9.14 648 10.6 816 8.71 984 6.46 1152 6.57 1320 3.41 1488 2.64 1656 1.93 1824 1.29 1992 1.13 2160 0.821 2328 0.763 2496 0.457 2664 0.562

[00319] Nalmefene time vs concentration data for compound 24 dose at 165 mg/kg are shown in Table 42.

Table 42 Time (h) Average concentration
nalmefene (ng/ml) 0.250 3.48 1.00 4.90 2.00 5.23 4.00 6.41 8.00 7.24 24.0 8.44 48.0 8.24 72.0 7.80 144 9.93 312 12.8 480 18.0 648 16.6 816 15.3 984 9.14 1152 5.62 1320 5.44 1488 3.62 1656 3.90 1824 2.69 1992 1.81 2160 1.28 2328 1.34 2496 0.886 2664 0.591

[00320] Naltrexone time vs. concentration data for Compound 29 (400 mg/ml in sesame oil) at 165 mg/kg are shown in Table 43a.

Table 43a Time (h) Average concentration
naltrexone (ng/ml) 0.250 13.8 1.00 30.4 2.00 39.6 4.00 53.2 8.00 53.1 24.0 46.2 48.0 34.3 72.0 30.4 144 39.6 312 28.4 480 13.1 648 6.00 816 4.56 984 3.88 1152 3.71 1320 2.71 1488 2.70 1656 2.45 1824 2.25 1992 1.92

[00321] Naltrexone time vs. concentration data for Compound 29 (300 mg/mL in cottonseed oil, n=2) at 165 mg/kg are shown in Table 43b.

Table 43b Time (h) Average concentration
naltrexone (ng/ml) 0.250 29.0 1.00 40.1 2.00 69.3 4.00 91.4 8.00 90.5 24.0 92.1 48.0 66.9 72.0 54.7

Example 2 Pharmacokinetic Studies in Dogs

Target

[00322] The purpose of this study was to determine the pharmacokinetic parameters of the test compounds in plasma after deep intramuscular administration to dogs of the Beagle breed (n=3, unless otherwise indicated).

Acclimatization/Quarantine

[00323] The general health of the animals was assessed and the animals were acclimatized for at least 5 days prior to use in the study.

Animal keeping

Animals were kept in pairs during acclimatization and kept separately during the study. The room(s) were monitored and controlled for relative humidity (target average range 40% to 70%, and any overshoot of this range lasting more than 3 hours documented as a deviation) and temperature (target average range 18°C). to 26°C, and any deviation outside this range was documented as a deviation) and air changes were carried out 10 to 20 times per hour. The room was maintained on a 12-hour cycle of alternating light and dark, except in cases where violations of this cycle were caused by activities within the study. Animals received food twice a day. Dogs of this breed receive approximately 220 g of certified dog food per day (Beijing Keao Xieli Feed Co., Ltd. Beijing, P. R. China). This amount of feed may be changed as necessary, depending on the feed intake of the group of animals or individual animals, changes in the body weight of the group of animals or an individual animal, or changes in the certified diet. All animals had unlimited access to reverse osmosis (RO) water. The reverse osmosis water was analyzed every three months, and each batch of feed was analyzed each time before use. Animals were provided with enriching toys.

Animal body weight and clinical observation

[00324] Body weight was determined prior to selection for use in the study and on the day of dosing. Body weight was monitored every week.

Extensive clinical observation, including observation of behavior and activity, reflection, respiration, skin and coat condition, facial expressions, genitourinary system, and the appearance of significant damage to other systems, was performed on the day of dose and each time samples were taken.

Dose Administration

[00325] The dose of the drug (at a concentration of 400 mg base equivalents / ml in sesame oil + 1% benzyl alcohol, unless otherwise indicated) was administered by deep intramuscular injection (unless otherwise indicated). Vehicle was also administered to each animal on the contralateral side at the start of the study by deep intramuscular injection (unless otherwise indicated). Animals were injected intravenously with sleeping pills, propofol at a dose of 6 mg/kg. After sedation, hair was carefully removed around the injection site, and the area was gently cleansed. Care was taken not to irritate the skin during shaving and cleaning the injection site. The dogs were then given a deep IM (intramuscular) injection by inserting the needle at least 2.5 cm from the surface into the central part of the quadriceps or biceps femoris muscle and guiding it towards the thigh. If the needle touched the thigh, the needle was simply pushed back a little and then the injection was made. The volume of the dose was determined depending on the body weight of the animal, which was measured on the morning of the day of administration. With repeated administration, the injection site may be changed to minimize tissue damage.

Table 44 Compound Nominal IM dose (mg/kg) Carrier Half-life t _1/2 (h) Half-life in the final phase t _1/2 (h) AUC _0-inf (ng.h/ml) prodrugs AUC _0-inf (ng.h/ml) nalmefene prodrug nalmefene 6 thirty Sesame oil + 1% benzyl alcohol Data in progress 48 (shallow
injection) Sesame oil + 1% benzyl alcohol 135 277.9 1025 11464 48 (deep
injection) Sesame oil + 1% benzyl alcohol 54 552.6 2186 10619 96 Sesame oil + 1% benzyl alcohol Data in progress 15 thirty Sesame oil + 1% benzyl alcohol Data in progress 48 Sesame oil + 1% benzyl alcohol 518 4244.8 1060 9684 96 Sesame oil + 1% benzyl alcohol Data in progress 24 48 (shallow
injection) Cottonseed Oil + 1% Benzyl Alcohol 102 308.8 526 8500 48 (deep
injection) Cottonseed Oil + 1% Benzyl Alcohol 59.6 404.0 613 7226 45 thirty Cottonseed Oil + 1% Benzyl Alcohol Data in progress 48 Cottonseed Oil + 1% Benzyl Alcohol Data in progress 96 Cottonseed Oil + 1% Benzyl Alcohol Data in progress 7 48 Sesame oil + 1% benzyl alcohol Data in progress 8 48 Sesame oil + 1% benzyl alcohol Data in progress 1 48 Sesame oil + 1% benzyl alcohol Data in progress 3 48 Sesame oil + 1% benzyl alcohol 4 74.4 360 9084 4 48 Sesame oil + 1% benzyl alcohol Data in progress 5 48 Sesame oil + 1% benzyl alcohol 70.9 105.5 3869 8579 35 48 Sesame oil + 1% benzyl alcohol Data in progress

Sampling

[00326] Blood samples were taken from the peripheral vessel from immobilized animals (without euthanasia) at each of the time sampling points.

[00327] Approximately 0.8 ml of blood was drawn at each time point. All blood samples were placed in pre-chilled plastic microcentrifuge tubes containing sodium fluoride/potassium oxalate (20 µl, 160 mg/ml, NaF/KO=1/3), 5% PMSF (100 mM in ethanol) as a stabilizer and EDTA- K ₂ (16 μl, 0.5 M) as an anticoagulant, which were kept in liquid ice until centrifugation.

Each blood sample obtained was kept in liquid ice prior to centrifugation.

Getting Plasma

[00328] Each blood sample obtained was centrifuged for 4 minutes at 4°C and 10,000 rpm to collect plasma. Plasma was collected and transferred to a pre-labeled PP tube placed in dry ice at each time point, and immediately precipitated using AN at a ratio of 1:4 (plasma:AN). Centrifuged again (10 minutes, 12,000 rpm) and the supernatant was collected.

After sampling was completed and prior to bioassay, the entire supernatant was stored at approximately -80°C.

Bioanalytical methodology and sample analysis

[00329] LC-MS/MS techniques have been developed to quantify a test compound in a biological environment. This method used a calibration curve with 8 non-zero calibration standards including LLOQ (0.05 ng/ml). Sample analysis was performed simultaneously with a set of calibration standards and two sets of QC samples using this LC-MS/MS method.

Data analysis

[00330] Plasma concentration versus time data was analyzed by non-compartmental analysis using Phoenix WinNonlin 6.3 software. Received the values of C _max , T _max , T _1/2 , AUC _(0-t) , AUC _(0-inf) , MRT _(0-t) , MRT _(0-inf) and graphs of plasma concentration versus time.

Nalmefene time vs. concentration data for a 30 mg/kg dose of Compound 6 are shown in Table 45.

Table 45* Time (h) Average concentration nalmefene (ng/ml) 0.25 8.07 1 11.50 2 7.75 4 4.33 8 3.41 24 10.14 48 10.82 72 12.15

*Research ongoing.

Nalmefene time vs. concentration data for a 48 mg/kg dose of Compound 6 (shallow IM injection) are shown in Table 46.

Table 46 Time (h) Average concentration nalmefene (ng/ml) 0.25 4.75 1 5.03 2 3.98 4 5.26 8 5.40 34 10.25 48 14.50 72 21.40 196 26.80 336 15.37 504 8.34 672 5.15 840 2.18 1008 1.82 1176 1.53 1344 1.07 1512 0.53

Nalmefene time vs. concentration data for the 48 mg/kg dose of Compound 6 (deep IM injection; repeated administration to dogs after the injection described in Table 46) are shown in Table 47a.

Table 47a* Time (h) Average concentration nalmefene (ng/ml) 0.25 8.99 1 18.97 2 13.23 4 9.70 8 11.08 24 12.91 48 15.83 72 19.93 168 21.93 336 12.00 504 5.59 672 4.31 840 2.24 1008 2.07 1176 1.86 1344 1.26 1512 1.11 2424 0.37

*Research ongoing.

Time vs concentration data for nalmefene for a 48 mg/kg dose of Compound 6 (deep IM injection; single dose administered to dogs not previously used in the study, n=2) are shown in Table 47b.

Table 47b* Time (h) Average concentration nalmefene (ng/ml) 0.25 17.3 1 17.0 2 13.2 4 9.99 8 7.73 24 12.3 48 17.6 72 25.0

*Research ongoing.

Nalmefene time vs. concentration data for a 96 mg/kg dose of Compound 6 are shown in Table 48.

Table 48* Time (h) Average concentration nalmefene (ng/ml) 0.25 13.59 1 24.17 2 14.11 4 12.56 8 6.84 24 17.80 48 22.13 72 33.97

*Research ongoing.

Nalmefene time vs. concentration data for a 30 mg/kg dose of Compound 15 are shown in Table 49.

Table 49* Time (h) Average concentration nalmefene (ng/ml) 0.25 5.74 1 10.04 2 5.49 4 2.84 8 0.67 24 0.41 48 0.56 72 0.78

*Research ongoing.

Data "time vs concentration of nalmefene" for the dose of compound 15 48 mg/kg are shown in Table 50a.

Table 50a* Time (h) Average concentration nalmefene (ng/ml) 0.25 6.74 1 15.67 2 10.28 4 4.23 8 1.19 24 1.65 48 1.95 72 3.14 168 2.92 336 3.94 504 2.71 672 1.96 840 1.90 1008 2.10 1176 1.70 1344 2.40 1512 2.44 2064 1.75 2232 1.58 2400 1.72 2568 1.46 2736 2.06

*Research ongoing.

Data "time vs concentration of nalmefene" for the dose of compound 15 48 mg/kg (repetition of the study described in Table 50a) are shown in Table 50b.

Table 50b* Time (h) Average concentration nalmefene (ng/ml) 0.25 7.55 1 12.3 2 10.3 4 4.48 8 1.55 24 0.965 48 1.53 72 1.64

*Research ongoing.

Nalmefene time vs. concentration data for a 96 mg/kg dose of Compound 15 are shown in Table 51.

Table 51* Time (h) Average concentration nalmefene (ng/ml) 0.25 18.63 1 24.13 2 16.83 4 9.81 8 2.84 24 2.38 48 2.65 72 2.98

*Research ongoing.

Nalmefene time vs. concentration data for a 48 mg/kg dose of Compound 18 are shown in Table 52.

Table 52 Time (h) Average concentration nalmefene (ng/ml) 0.25 3.49 1 5.44 2 5.18 4 6.62 8 9.77 24 14.40 48 14.24 72 16.13

Nalmefene time vs. concentration data for a 48 mg/kg dose of Compound 24 (shallow IM injection) are shown in Table 53.

Table 53 Time (h) Average concentration nalmefene (ng/ml) 0.25 1.33 1 1.62 2 2.17 4 4.70 8 4.71 24 8.34 48 10.93 72 15.83 168 24.07 336 11.29 504 5.13 672 2.42 840 1.59 1008 1.26 1176 0.86 1344 0.56 1512 0.50

Time vs. concentration data for nalmefene for a 48 mg/kg dose of Compound 24 (deep IM injection) are shown in Table 54.

Table 54 Time (h) Average concentration nalmefene (ng/ml) 0.25 1.64 1 2.03 2 2.11 4 2.23 8 3.47 24 5.82 48 11.60 72 15.90 168 14.63 336 9.96 504 5.85 672 2.47 840 1.99 1008 1.51 1176 1.07 1344 0.85 1512 0.54

Nalmefene time vs. concentration data for a 30 mg/kg dose of Compound 45 are shown in Table 55.

Table 55* Time (h) Average concentration nalmefene (ng/ml) 0.25 2.82 1 4.04 2 3.24 4 2.75 8 2.61 24 3.00 48 5.31 72 4.72

*Research ongoing.

Nalmefene time vs. concentration data for a 48 mg/kg dose of Compound 45 are shown in Table 56.

Table 56* Time (h) Average concentration nalmefene (ng/ml) 0.25 1.22 1 2.23 2 1.77 4 1.75 8 1.95 24 3.05 48 10.00 72 10.05 168 18.87 336 14.83 504 7.43 672 3.24

*Research ongoing.

Nalmefene time vs. concentration data for a 96 mg/kg dose of Compound 45 are shown in Table 57.

Table 57* Time (h) Average concentration nalmefene (ng/ml) 0.25 2.93 1 7.71 2 6.64 4 4.86 8 3.66 24 7.14 48 11.30 72 9.32

*Research ongoing.

Data "time vs concentration of naltrexone" for the dose of compound 7 24 mg/kg are shown in Table 58.

Table 58* Time (h) Average concentration naltrexone (ng/ml) 0.25 3.61 1 5.25 2 2.52 4 0.82 8 0.29 24 0.61 48 0.61 72 0.67

*Research ongoing.

Data "time vs concentration of naltrexone" for the dose of compound 7 48 mg/kg (n=2) are shown in Table 59.

Table 59* Time (h) Average concentration naltrexone (ng/ml) 0.25 18.23 1 18.13 2 9.67 4 3.35 8 1.33 24 1.43 48 2.30 72 2.41 168 1.85 336 1.09 504 1.35 672 1.27

*Research ongoing.

Data "time vs concentration of naltrexone" for the dose of compound 8 48 mg/kg are shown in Table 60.

Table 60* Time (h) Average concentration naltrexone (ng/ml) 0.25 103.20 1 159.67 2 52.33 4 9.74 8 5.03 24 3.46 48 4.06 72 4.87 168 7.28 336 3.91 504 9.72 672 4.59

*Research ongoing.

Nalmefene time vs. concentration data for a 48 mg/kg dose of Compound 1 are shown in Table 61.

Table 61* Time (h) Average concentration nalmefene (ng/ml) 0.25 1.90 1 4.24 2 2.26 4 0.92 8 0.57 24 0.87 48 1.41 72 1.84 168 2.57 336 1.78 504 1.49 672 1.21

*Research ongoing.

Nalmefene time vs. concentration data for a 48 mg/kg dose of Compound 3 are shown in Table 62.

Table 62 Time (h) Average concentration nalmefene (ng/ml) 0.25 2.54 1 4.48 2 5.37 4 9.26 8 13.37 24 25.43 48 37.57 72 62.60 168 27.77 336 4.50 504 0.97 672 0.34

Data "time vs concentration of naltrexone" for the dose of compound 4 48 mg/kg are shown in Table 63.

Table 63 Time (h) Average concentration naltrexone (ng/ml) 0.25 21.90 1 38.67 2 41.17 4 74.47 8 78.07 24 53.23 48 52.93 72 70.03 168 3.76 336 ND** 504 ND 672 ND

**ND = not found.

Data "time vs concentration of naltrexone" for the dose of compound 5 48 mg/kg (n=2) are shown in Table 64.

Table 64 Time (h) Average concentration nalmefene (ng/ml) 0.25 1.12 1 2.75 2 3.55 4 5.49 8 6.67 24 17.40 48 26.95 72 30.65 168 27.85 336 11.00 504 2.07 672 1.06

Data "time vs concentration of naltrexone" for the dose of compound 35 48 mg/kg are shown in Table 65.

Table 65 Time (h) Average concentration nalmefene (ng/ml) 0.25 9.57 1 19.07 2 14.50 4 18.80 8 25.80 24 103.87 48 174.67 72 237.00

Clinical data for dogs treated with Compound 6 at 30 mg/kg are shown in Table 66.

Time point D1501 D1502 D1503 L*-carrier R*-TA** L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

*L - injection in the left leg; R - injection in the right leg;

**TA - therapeutic agent.

Clinical data for dogs treated with Compound 6 at 48 mg/kg (shallow IM injection) are shown in Table 67.

Time point D1001 D1002 D1003 R-TA L-carrier R-TA L-carrier R-TA L-carrier 8 ocloc'k Without pronounced observable clinical manifestations NA Without pronounced observable clinical manifestations NA Without pronounced observable clinical manifestations NA 72 hours Without pronounced observable clinical manifestations NA Without pronounced observable clinical manifestations NA Slight swelling NA Day 5 Without pronounced observable clinical manifestations NA Slight swelling NA More pronounced swelling NA Day 7 Without pronounced observable clinical manifestations NA Slight swelling NA More pronounced swelling NA Day 8 Slight swelling, induration NA Slight swelling, induration NA More pronounced swelling, induration NA Day 9 No change from Day 8 NA No change since Day 8 NA No change since Day 8 NA Day 12 No significant swelling and induration, no inflammation, no pain when touched NA No significant swelling and induration, no inflammation, no pain when touched NA Significant swelling and induration, no inflammation, no pain when touched NA Day 16 No change from Day 15 Without pronounced observable clinical manifestations No change since Day 15 Without pronounced observable clinical manifestations No change from Day 15 Without pronounced observable clinical manifestations Day 17 More pronounced induration than at the site of vehicle injection Without pronounced observable clinical manifestations More pronounced induration than at the site of vehicle injection Without pronounced observable clinical manifestations More pronounced induration than at the site of vehicle injection Without pronounced observable clinical manifestations Day 18 No change from Day 17 Without pronounced observable clinical manifestations No change from Day 17 Without pronounced observable clinical manifestations No change from Day 17 Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 22 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 30 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 42 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 49 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 56 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 63 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 6 at 48 mg/kg (deep IM injection; repeated administration to dogs after the injection described in Table 67) are shown in Table 68a.

Time point D1001 D1002 D1003 L-TA R-carrier L-TA R-carrier L-TA R-carrier 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 42 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 49 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 56 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 63 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 101 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 6 at 48 mg/kg (deep IM injection; single dose administered to dogs not previously used in the study, n=2) are shown in Table 68b.

Time point D1001 D1002 L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Vomiting: Approximately 20 g chyme 1 hour post dose Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 6 at 96 mg/kg are shown in Table 69.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 15 at 30 mg/kg are shown in Table 70.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Extensive area of red spots on the outside and inside of the leg / red spots on the outside at the stage of scab removal Extensive area of red spots on the outside and inside of the leg / red spots on the outside at the stage of scab removal Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 18 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Extensive area of red spots on the outer side of the leg / red spots on the outer side at the stage of removal of the crust and formation of a scab Extensive area of red spots on the outer side of the leg / red spots on the outer side at the stage of removal of the crust and formation of a scab Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 20 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Regression of red spots on the outside of the leg and removal of the crust Regression of red spots on the outside of the leg,
eschar formation / eschar measuring 1.5 cm * 1.5 cm on the inside of the right leg Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Removing the crust on the outside of the left leg Eschar formation on the outside of the leg / 1.5 cm * 1.5 cm eschar on the inside of the right leg Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 25 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Removing the crust on the outside of the left leg Removing the crust on the outside of the right leg Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 15 at 48 mg/kg are shown in Table 71a.

Time point D1501 D1502 D1503 L-TA R-carrier L-TA R-carrier L-TA R-carrier until Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling all over the leg Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 8 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling in the thigh / swelling in the lower leg Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 9 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling in the thigh / swelling in the lower leg / skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 10 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling in the thigh/skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 11 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 9 * 7 cm in the thigh / skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 12 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 9 * 7 cm in the thigh / skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 13 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Slight swelling in the thigh/skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Slight swelling in the thigh/skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 15 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Slight swelling in the thigh/skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 16 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Slight swelling in the thigh/skin rash in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 17 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 42 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 49 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 56 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 63 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 86 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 93 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 100 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 107 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 114 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 15 at 48 mg/kg are shown in Table 71b.

Time point D1001 D1002 D1003 L-carrier R-TA L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 15 at 96 mg/kg are shown in Table 72.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 8*4 cm Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 18 at 48 mg/kg are shown in Table 73.

Time point D1501 D1502 D1502 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Approximately 10 ml loose stools 8 hours after administration 24 hours Without pronounced observable clinical manifestations Thigh induration 18*10 cm/ inner thigh swelling 6*9 cm/ body temperature: 39.1°C Red spots in the thigh area Swelling 15*12 cm in the thigh / swelling 8*11 on the inner thigh with redness and a feeling of heat / swelling and severe redness on the back of the lower leg / body temperature: 40.1°C Without pronounced observable clinical manifestations Induration 15*13 cm of the entire leg with redness and a feeling of heat/ Induration on the inner thigh 9*11 cm with redness and a feeling of heat/body temperature: 39.9°C 48 hours Swelling 3*6cm Induration in the thigh area 18*10 cm/ swelling on the inner surface of the thigh 8*9 cm/ red spots at 1 o'clock of the conventional dial/ body temperature: 39.1°C Red spots in the thigh area Swelling 15*12 cm in the thigh / swelling 8*11 on the inner thigh with redness and a feeling of heat / swelling and darkening on the back of the leg / body temperature: 40.0 ° C Without pronounced observable clinical manifestations Induration 15*13 cm of the entire leg with redness and a feeling of heat/ Induration on the inner thigh 9*11 cm with redness and a feeling of heat/body temperature: 39.9°C 72 hours Swelling 3*6cm Induration in the thigh area 18*10 cm/ swelling on the inner surface of the thigh 8*9 cm/ red spots at 1 o'clock of the conventional dial/ body temperature: 39.1°C/ loose stools/ lack of appetite Red spots in the thigh area Swelling 15 * 12 cm in the thigh / swelling 8 * 11 on the inner thigh with redness and a feeling of heat / swelling and darkening on the back of the lower leg / body temperature: 38.7 ° C / ulcerated scabs 2 * 4 on the inner thigh / loose stools / lack of appetite Without pronounced observable clinical manifestations Induration 15*13 cm of the whole leg with redness and a feeling of heat / Induration on the inner thigh 9*11 cm with redness and a feeling of heat / body temperature: 39.4°C / loose stools / lack of appetite 96 hours Swelling 3*6cm Lameness/ Severe induration of the entire leg/ Ankle edema/ Red spots at 1 o’clock/ Body temperature: 39.1°C/ Loose stools/ Loss of appetite/ Weight loss from 6.71 kg to 6.0 kg/ Bloody chair Slight swelling / red spots in the thigh area Lameness / induration of the entire leg / edema in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 1.5 * 1.5 cm in the thigh and 6 * 2 cm in the lower leg with red secretion / body temperature: 39.0°C / loose stools / lack of appetite / weight loss from 7.76 kg to 6.86 kg / bloody stools Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / body temperature: 39.4 ° C / loose stools / lack of appetite / weight loss from 6.52 kg to 6.17 kg / bloody stools 120 hours Swelling 3*6cm Lameness/ Severe induration of the whole leg/ Swelling in the ankle/ Red spots at 1 o'clock on the dial/ Body temperature: 39.8°C/ Loose stools/ Loss of appetite Slight swelling / red spots in the thigh area Lameness / induration of the entire leg / edema in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 1.5 * 1.5 cm in the thigh and 6 * 2 cm in the lower leg with red secretion / body temperature: 39.1°C / loose stools / lack of appetite Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / body temperature: 39.5°C / loose stools 144 hours Swelling 3*6cm Lameness/ severe induration of the whole leg/ swelling in the ankle area/ red spots at 1 o'clock of the conventional dial/ induration on the inner thigh of a rich color and with a feeling of heat/ body temperature: 39.3°C/ loose stools/ lack of appetite/ 6.18 kg Slight swelling / red spots in the thigh area Lameness / induration of the entire leg / edema in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 1.5 * 1.5 cm in the thigh and 6 * 2 cm in the lower leg with red secretion / body temperature: 39.2°C / loose stools / lack of appetite / 6.92 kg Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / body temperature: 39.5°C / loose stools / 6.15 kg 168 hours Swelling 3*6cm Lameness/severe induration of the entire leg/swelling in the ankle/red spots at 1 o'clock/induration on the inner thigh of a rich color and with a feeling of heat/ body temperature: 39.2°C/ loose stools/ loss of appetite/ 6.13 kg Slight swelling / red spots in the thigh area Lameness / induration of the entire leg / edema in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 1.5 * 1.5 cm in the thigh and 6 * 2 cm in the lower leg with red secretion / body temperature: 39.6°C / loose stools / lack of appetite / 7.01 kg Without pronounced observable clinical manifestations Induration of the whole leg with redness and a feeling of heat / ulceration in the thigh 0.5 * 0.5 cm / body temperature: 38.7 ° C / liquefied stool / 5.93 kg Day 8 Swelling 3*6cm/ Lameness/severe induration of the entire leg/swelling of the ankle/ulceration with red and thick secretions in the thigh/induration of the inner thigh, intense color and with a feeling of heat/body temperature: 39.4°C/thin stools/lack of appetite/6 .11 kg Slight swelling / red spots in the thigh area Lameness / induration of the entire leg / edema in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 3 * 1.5 cm in the thigh and 6 * 4 cm in the lower leg with red secretion and muscle exposure / body temperature: 39.5°C / loose stools / lack of appetite / 6.76 kg Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / ulceration in the thigh area 0.5 * 0.5 cm with redness and a feeling of heat / Induration on the anterior surface of the leg and fluctuation of red and thick fluid on the back of the leg / body temperature: 38.7 ° C/ loose stool/ 5.98 kg Day 9 Swelling 3*6cm/ Lameness/ Severe induration of the entire leg/ Swelling of the ankle/ 3 parts of ulceration with yellow and thick secretions in the thigh/ Induration of the inner thigh, intense color and feeling hot/ Body temperature: 38.7°C/ Loose stools/ Loss of appetite / 6.09 kg Slight swelling / red spots in the thigh area Lameness / induration of the entire leg / edema in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 3 * 1.5 cm in the thigh and 6 * 4 cm in the lower leg with yellow secretion and muscle exposure / body temperature: 38.5°C / loose stools / lack of appetite / 6.79 kg Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / ulceration in the thigh area 0.5 * 0.5 cm with redness and a feeling of heat / induration on the anterior surface of the leg and fluctuation of red and thick fluid on the back of the leg / body temperature: 38.8 ° C/ loose stool/ 6.15 kg Day 10 Swelling 3*6cm/ Lameness/severe induration of the entire leg/swelling of the ankle/3 parts of ulceration with yellow and thick secretion in the thigh area, one part of ulceration with pink and thick secretion on the inner thigh/induration on the inner thigh of a rich color and with a feeling of heat/temperature body: 38.3°C / loose stools / no appetite / 6.05 kg Without pronounced observable clinical manifestations Induration of the entire leg / swelling in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 3 * 1.5 cm in the thigh and 6 * 4 cm in the lower leg with yellow secretion and muscle exposure / temperature body: 38.8°C / loose stools / no appetite / 6.68 kg Without pronounced observable clinical manifestations Induration of the whole leg with redness and a feeling of heat / ulceration in the thigh area 0.5 * 0.5 cm with redness and a feeling of heat and the appearance of scabs / induration on the anterior surface of the lower leg and fluctuation of red and thick fluid on the back of the lower leg / body temperature: 38 .7°C/ loose stool/ 6.13 kg Day 11 Without pronounced observable clinical manifestations Lameness/severe induration of the entire leg/swelling of the ankle/3 parts of ulceration with yellow and thick secretion in the thigh area, one part of ulceration with pink and thick secretion on the inner thigh/induration on the inner thigh of a rich color and with a feeling of heat/temperature body: 38.5°C / loose stools / no appetite / 5.99 kg Without pronounced observable clinical manifestations Induration of the entire leg / swelling in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 3 * 1.5 cm in the thigh and 6 * 4 cm in the lower leg with yellow secretion and muscle exposure / temperature body: 38.9°C / loose stools / no appetite / 6.83 kg Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / ulceration in the thigh area 0.5 * 0.5 cm with redness and a feeling of heat and the appearance of scabs / induration on the anterior surface of the leg and fluctuation of red and thick fluid on the back of the leg / body temperature: 38 .7°C/ loose stool/ 6.13 kg Day 12 Without pronounced observable clinical manifestations Lameness/ Severe induration of the entire leg/ Swelling in the ankle/ 3 parts of ulceration with yellow and thick secretions in the thigh area, one part of ulceration on the inner thigh/ Induration on the inner thigh of deep color and with a feeling of heat/ loose stools/ lack of appetite Without pronounced observable clinical manifestations Induration of the entire leg / swelling in the distal leg / induration and darkening in the proximal leg / necrosis (darkening and muscle atrophy) 3 * 1.5 cm in the thigh and 6 * 4 cm in the lower leg with yellow secretions and muscle exposure / liquefied stool / lack of appetite / Without pronounced observable clinical manifestations Induration of the entire leg with redness and a feeling of heat / ulceration in the thigh 0.5 * 0.5 cm with redness and a feeling of heat and the appearance of scabs / loose stools Day 13 Without pronounced observable clinical manifestations Lameness / swelling on the inner thigh purple / swelling in the ankle / 3 parts of ulceration with yellow and thick secretion in the thigh, one part of ulceration with pink and thick secretion on the inner thigh / 6.06 kg Without pronounced observable clinical manifestations Induration of the entire leg/ necrosis (darkening and muscle atrophy) 1.5*1.5 cm in the thigh and 5*1.5 cm in the lower leg with muscle exposure/ 6.92 kg Without pronounced observable clinical manifestations Whole leg induration / ulceration in the thigh area, covered with scabs / 6.45 kg Day 14 Without pronounced observable clinical manifestations Lameness / swelling on the inner surface of the thigh purple / swelling in the ankle / 3 parts of ulceration with yellow and thick secretion in the thigh area, one part of ulceration with pink and thick secretion on the inner thigh / 6.01 kg Without pronounced observable clinical manifestations Whole leg induration / necrosis (darkening and muscle atrophy) 1.5*1.5 cm in the thigh and 5*1.5 cm in the lower leg with muscle exposure / 6.93 kg Without pronounced observable clinical manifestations Whole leg induration / ulceration in the thigh area, covered with scabs / 6.38 kg Day 15 Without pronounced observable clinical manifestations Lameness / swelling on the inner thigh purple / swelling in the ankle / 3 parts ulceration with yellow and thick secretion in the thigh, one part ulceration with pink and thick secretion on the inner thigh Without pronounced observable clinical manifestations Whole leg induration/necrosis (darkening and muscle atrophy) 1.5*1.5 cm in the thigh and 5*1.5 cm in the lower leg with muscle exposure Without pronounced observable clinical manifestations Whole leg induration/scabbing ulceration in thigh

Clinical data for dogs treated with Compound 24 at 48 mg/kg (shallow IM injection) are shown in Table 74.

Time point D1001 D1002 D1003 L-carrier R-TA L-carrier R-TA L-carrier R-TA 2 hours Without pronounced observable clinical manifestations Swelling Without pronounced observable clinical manifestations Swelling Without pronounced observable clinical manifestations Swelling 8 ocloc'k Without pronounced observable clinical manifestations More pronounced swelling Without pronounced observable clinical manifestations More pronounced swelling Without pronounced observable clinical manifestations More pronounced swelling 24 hours Without pronounced observable clinical manifestations More pronounced swelling Without pronounced observable clinical manifestations More pronounced swelling and eschar along the edge of the swelling area Without pronounced observable clinical manifestations More pronounced swelling 48 hours Without pronounced observable clinical manifestations More pronounced swelling Without pronounced observable clinical manifestations More pronounced swelling and eschar along the edge of the swelling area Without pronounced observable clinical manifestations More pronounced swelling 72 hours Without pronounced observable clinical manifestations More pronounced swelling Without pronounced observable clinical manifestations More pronounced swelling and eschar along the edge of the swelling area Without pronounced observable clinical manifestations More pronounced swelling 96 hours Without pronounced observable clinical manifestations More pronounced swelling Without pronounced observable clinical manifestations More pronounced swelling and eschar along the edge of the swelling area Without pronounced observable clinical manifestations More pronounced swelling 120 hours Without pronounced observable clinical manifestations More pronounced swelling with induration Without pronounced observable clinical manifestations More pronounced swelling with induration and eschar along the edge of the swelling zone Without pronounced observable clinical manifestations More pronounced swelling with induration 144 hours 0.1 * 0.2 cm / several areas of redness More pronounced swelling with induration 2 yellow scabs 0.8*0.5 cm each, one yellow scab with a red edge More pronounced swelling with induration and eschar along the edge of the swelling zone Without pronounced observable clinical manifestations More pronounced swelling with induration 178 hours 4 yellow scabs 0.1*0.2 cm each / superficial damage Induration / slight redness / size: 5*8 cm / depth: 0.5-0.8 cm 3 yellow scabs 0.2*0.2 cm each / superficial lesion (one scab covers two areas of redness) Induration / size: 5*8 cm / depth: 0.5-0.8 cm; 0.3*0.3 cm/ red scab Without pronounced observable clinical manifestations Induration / size: 5*5 cm / depth: 0.5-1 cm 192 hours 4 yellow scabs 0.1*0.2 cm each / superficial damage Induration / slight redness / size: 5*8 cm / depth: 0.5-0.8 cm 2 yellow scabs 0.2 * 0.2 each; 2 yellow scabs 0.3*0.5 cm each size: 0.3*0.1 cm/ red scab induration/ 5*8 cm/ depth: 0.5-0.8 cm; 0.3*0.3 cm/ red scab Without pronounced observable clinical manifestations Induration / size: 5*5 cm / depth: 0.5-1 cm 216 hours 4 yellow scabs 0.1*0.2 cm each / superficial damage / reduction of damage Induration / slight redness / size: 4*8 cm / depth: 0.5-0.8 cm Yellow eschar / superficial damage / light damage / damage reduction size: 0.2 * 0.1 cm / red scab / damage reduction; Induration / 4*8 cm / depth: 0.5-0.8 cm; restoration of skin color with the appearance of two yellow scabs / 1 * 1 cm, 2 * 2 cm Without pronounced observable clinical manifestations Induration / size: 5*8 cm / depth: 0.5-1 cm 240 hours 4 yellow scabs 0.1*0.2 cm each / superficial damage / reduction of damage Induration / size: 7*7 cm Yellow eschar/superficial injury/slight injury/damage reduction size: 0.2 * 0.1 cm / red scab / damage reduction; Induration / 4 * 8 cm /; restoration of skin color with the appearance of two yellow scabs / 1 * 1 cm, 2 * 2 cm Without pronounced observable clinical manifestations Induration / size: 5*7 cm 264 hours 4 yellow scabs 0.1 * 0.1 cm each / superficial damage / reduction of damage Induration / size: 7 * 7 cm / Yellow eschar / superficial damage / light damage / damage reduction size: 0.1*0.1 cm / red scab / damage reduction; Induration / 7 * 7 cm /; two yellow scabs / 1 * 1 cm, 2 * 2 cm / superficial damage Without pronounced observable clinical manifestations Induration / size: 9 * 3.5 cm / 288 hours Yellow eschar / superficial damage / slight damage / reduction of damage to a red spot Induration / size: 7 * 7 cm / Yellow scab / disappearance of damage, skin restoration; Induration / 7 * 8 cm /; two yellow scabs / 1 * 1 cm, 2 * 2 cm / superficial damage No significant pathology Induration / size: 9 * 5.5 cm / 312 hours Yellow eschar/superficial injury/slight injury/damage reduction; 1 zone with red rash / 0.2*0.2 cm Induration / slight redness / size: 10 * 6 cm (largest size * smallest size, in the form of "short boot") / depth: 0.5-0.8 cm; skin rash in the groin and axilla / moderate damage Skin repair / skin repair at the site of the scab; new yellow scab/ superficial damage/ 1.5*2 cm Induration / slight redness / size: 10 * 6 cm / depth: 0.5-0.8 cm; 3 yellow scabs / moderate damage; restoration of skin color / redness / 3 * 2 cm / small red spot Without pronounced observable clinical manifestations Induration / size: 9*5 cm / depth: 0.5-1.5 cm 336 hours Without pronounced observable clinical manifestations Induration/ slight redness/ size: 10*6 cm (largest size*smallest size, in the form of a "short boot")/ skin rash in the groin area Yellow scab / superficial damage / 2*2 cm Induration / size: 10*6 cm; 5 yellow scabs (2*1 cm, 2*2 cm, 1*1 cm, 3*2 cm, 2*0.5 cm) on the right and one scab 2*2 on the left / moderate ulcer Without pronounced observable clinical manifestations Induration / size: 9*5 cm 360 hours Without pronounced observable clinical manifestations Induration / slight redness / size: 10*6 cm (largest size * smallest size, in the form of "short boot") / skin rash in the groin and armpit Yellow eschar/ superficial damage/ 2*2 cm/ alopecia Induration / size: 10*6 cm; 5 yellow scabs (2*1 cm, 2*2 cm, 1*1 cm, 3*2 cm, 2*0.5 cm) on the right and one scab 2*2 on the left / alopecia Without pronounced observable clinical manifestations Induration / size: 9*5 cm 384 hours Without pronounced observable clinical manifestations Induration / slight redness / size: 9.5*6 cm (largest size * smallest size, in the form of a "short boot") / skin rash in the groin and armpit / moderate damage Yellow eschar/ superficial damage/ 2*2 cm/ alopecia Induration, softening the edge of the induration zone / size: 10 * 7 cm; 4 yellow scabs (1*1 cm, 2*2 cm, 3*2 cm, 2*0.5 cm) on the right and one scab 2*2 on the left / alopecia Without pronounced observable clinical manifestations Induration / size: 9*5 cm 408 hours Without pronounced observable clinical manifestations Induration / slight redness / size: 10 * 7 cm (largest size * smallest size, in the form of "short boot") / skin rash in the groin Yellow eschar/ superficial damage/ 2*2 cm/ alopecia Induration, softening the edge of the induration zone / size: 10 * 7 cm; 4 yellow scabs (1*1 cm, 2*2 cm, 3*2 cm, 2*0.5 cm) on the right and one scab 2*2 on the left / alopecia Without pronounced observable clinical manifestations Induration / size: 11*6 cm 432 hours Without pronounced observable clinical manifestations Induration / slight redness / size: 10*6 cm (largest size * smallest size, in the form of "short boot") / skin rash in the groin and armpit Yellow eschar/ superficial damage/ 2*2 cm/ alopecia Induration, softening the edge of the induration zone / size: 10 * 6 cm; 3 yellow scabs (1*1 cm, 3*2 cm, 2*0.5 cm) on the right and one scab 2*2 on the left / alopecia Without pronounced observable clinical manifestations Induration / size: 10*5 cm 456 hours Without pronounced observable clinical manifestations Induration / slight redness / size: 10*6 cm (largest size * smallest size, in the form of "short boot") / skin rash in the groin and armpit Yellow eschar/ superficial damage/ 2*2 cm/ alopecia Induration, softening the edge of the induration zone / size: 11 * 7 cm (oval); 3 yellow scabs (1*1 cm, 3*2 cm, 2*0.5 cm) on the right and one scab 2*2 on the left / alopecia Without pronounced observable clinical manifestations Induration / size: 9*6 cm (oval) 480 hours Without pronounced observable clinical manifestations Induration / slight redness / size: 10 * 7 cm (largest size * smallest size, in the form of a "short boot") / skin rash in the groin Yellow eschar/ superficial damage/ 2*2 cm/ alopecia Induration, softening the edge of the induration zone / size: 11 * 7 cm (oval); 3 yellow scabs (1*1 cm, 3*2 and 2*0.5 cm) on the right and one scab 2*2 on the left / two red blisters / alopecia Without pronounced observable clinical manifestations Induration / size: 9*6 cm (oval) 504 hours Without pronounced observable clinical manifestations Decreased induration / 9*3 cm / induration near the right knee, depth: 0.2~0.5 / skin rash in the groin Alopecia Reducing induration, softening the edge of the induration zone / size: 9 * 6 cm; 2 yellow scabs (1 * 1 cm and 1.5 * 0.8 cm) on the right / two red blisters / alopecia Without pronounced observable clinical manifestations Induration reduction / size: 7*3 cm 528 hours Rash at site of vehicle injection Decreased induration / 9*3 cm / induration near the right knee, depth: 0.2~0.5 / skin rash in the groin and armpit Alopecia Reducing induration, softening the edge of the induration zone / size: 8 * 6 cm; 1 yellow scab (0.8 * 0.7 cm) on the right / two red blisters / alopecia Without pronounced observable clinical manifestations Induration Reduction/Size: 7*3cm 552 hours Rash gives rise to scabs Decreased induration / 8*3 cm / induration near the right knee, depth: 0.2~0.5 / skin rash in the groin and axilla / scab 0.8*0.6 cm Alopecia Reducing induration, softening the edge of the induration zone / size: 8 * 5 cm; 1 yellow scab (0.8 * 0.7 cm) on the right / two red blisters / alopecia Without pronounced observable clinical manifestations Induration reduction / size: 7*3 cm 576 hours Rash gives rise to scabs Decreased induration / 7*3 cm / induration near the right knee, depth: 0.2~0.5 / skin rash in the groin / scab 0.8*0.6 cm Alopecia Reducing induration, softening the edge of the induration zone / size: 7 * 5 cm; / two red blisters / alopecia Without pronounced observable clinical manifestations Induration reduction / size: 6*3 cm 600 hours Rash gives rise to scabs Decreased induration / 7*3 cm / induration near the right knee, depth: 0.2~0.5 / skin rash in the groin and armpit / scab 0.8*0.6 cm Alopecia Reducing induration, softening the edge of the induration zone / size: 7 * 5 cm; /two red blisters/alopecia Without pronounced observable clinical manifestations Induration reduction / size: 6*3 cm 624 hours Rash gives rise to scabs Decreased induration / 7*3 cm / induration near the right knee, depth: 0.2~0.5 / skin rash in the groin and armpit / scab 0.8*0.6 cm Alopecia Reducing induration, softening the edge of the induration zone / size: 7 * 5 cm; / two red blisters / alopecia Without pronounced observable clinical manifestations Induration Reduction/Size: 6*3cm 648 hours Rash gives rise to scabs Reducing induration / 7*3 cm/ skin rash in the groin and armpit Without pronounced observable clinical manifestations Reducing induration, softening the edge of the induration zone / size: 7 * 5 cm; Without pronounced observable clinical manifestations Decrease induration 672 hours Rash Reduction of induration / 5.5*3 cm/ skin rash in the groin and armpit Rash Reducing induration, softening the edge of the induration zone / size: 7 * 4 cm; Without pronounced observable clinical manifestations Swelling/6.5*3cm 696 hours Rash Reduction of induration / 5*3 cm/ skin rash in the groin and armpit Rash reduction Reducing induration, softening the edge of the induration zone / size: 6.5 * 4 cm; Without pronounced observable clinical manifestations Swelling/5.5*3cm 720 hours Rash Induration reduction / 4.5*3 cm / skin rash in the groin area Without pronounced observable clinical manifestations Reducing induration, softening the edge of the induration zone / size: 5 * 4 cm; Without pronounced observable clinical manifestations Swelling/5*3cm 744 hours Rash Reduction of induration / 4*3 cm/ skin rash in the groin and armpit Without pronounced observable clinical manifestations Reducing induration, softening the edge of the induration zone / size: 5 * 4 cm; Without pronounced observable clinical manifestations Swelling / 5 * 3 cm / induration in the center of the swelling zone 768 hours Rash Reduction of induration / 4*3 cm/ skin rash in the groin and armpit Without pronounced observable clinical manifestations Reducing induration, softening the edge of the induration zone / size: 5 * 4 cm; rash found in the stomach area Without pronounced observable clinical manifestations Swelling / 5 * 3 cm / induration in the center of the swelling zone 792 hours Rash Reduction of induration / 4*3 cm/ skin rash in the groin and armpit Without pronounced observable clinical manifestations Reducing induration, softening the edge of the induration zone / size: 5 * 4 cm; rash found in the stomach area Without pronounced observable clinical manifestations Swelling / 5 * 3 cm / induration in the center of the swelling zone 816 hours Rash Reduction of induration / 4*3 cm/ skin rash in the groin and armpit Without pronounced observable clinical manifestations Reducing induration, softening the edge of the induration zone / size: 5 * 3 cm; rash found in the stomach area Without pronounced observable clinical manifestations Swelling / 4 * 3 cm / induration in the center of the swelling zone 840 hours Rash Decreased induration / 4*3 cm / skin rash in the groin and armpit / body temperature: 38.5°C Swelling / redness / sensation of heat Whole leg swelling/redness/feeling hot/body temperature: 38.5°C Without pronounced observable clinical manifestations Swelling / 4*3 cm / rash in the groin / body temperature: 38.4°C 864 hours Rash Decreased induration / 4*3 cm / skin rash in the groin and axilla / body temperature: 38.7°C Swelling / redness / sensation of heat Swelling 5*3 cm in the thigh and swelling of the entire back of the lower leg / redness / sensation of heat / body temperature: 38.9 ° C Without pronounced observable clinical manifestations Swelling / 4*3 cm / rash in the groin / body temperature: 38.8°C 888 hours Rash Decreased induration / 4*3 cm / skin rash in the groin and axilla / body temperature: 38.0°C Swelling / redness / sensation of heat Swelling 5*3 cm in the thigh area and swelling of the entire back of the leg / redness / sensation of heat / body temperature: 38.5 ° C Without pronounced observable clinical manifestations Swelling / 4*3 cm / rash in the groin / body temperature: 38.6°C 912 hours Rash Decreased induration / 3*2 cm / skin rash in the groin and armpit / body temperature: 38.6°C Swelling / redness / sensation of heat Swelling 5*3 cm in the thigh and swelling of the entire back of the lower leg / redness / sensation of heat / body temperature: 38.9 ° C Without pronounced observable clinical manifestations Swelling / 4*3 cm / rash in the groin / body temperature: 38.6°C 936 hours Rash Decreased induration / 3*2 cm / skin rash in the groin and axilla / body temperature: 38.2°C Redness/feeling hot Swelling 3*2 cm in the thigh area and swelling of the entire back of the lower leg / redness / sensation of heat / body temperature: 38.9 ° C Without pronounced observable clinical manifestations Swelling / 3*2 cm / rash in the groin / body temperature: 38.6 ° C 960 hours Rash Decreased induration / 3*2 cm / skin rash in the groin and armpit / body temperature: 38.5°C Without pronounced observable clinical manifestations Swelling 3*2 cm in the thigh / body temperature: 39.0°C Without pronounced observable clinical manifestations Swelling / 3*2 cm / rash in the groin / body temperature: 38.8°C 984 hours Rash Decreased induration / 2*2 cm / skin rash in the groin and axilla / body temperature: 38.3°C Without pronounced observable clinical manifestations Swelling 3*2 cm in the thigh / body temperature: 39.2°C Without pronounced observable clinical manifestations Swelling / 3*2 cm / rash in the groin / body temperature: 38.9 ° C 1008 hours Rash Decreased induration / 2*2 cm / skin rash in the groin and armpit / body temperature: 38.6°C Without pronounced observable clinical manifestations Swelling 3*2 cm in the thigh / body temperature: 38.9°C Without pronounced observable clinical manifestations Swelling / 3*2 cm / rash in the groin / body temperature: 38.6°C 1032 hours Rash Skin rash in the groin and armpit Without pronounced observable clinical manifestations Swelling 3*2 cm in the thigh area Without pronounced observable clinical manifestations Swelling / 2 * 2 cm / rash in the groin 1056 hours Rash Skin rash in the groin and armpit / body temperature: 38.3°C Without pronounced observable clinical manifestations Swelling 3*2 cm in the thigh / body temperature: 38.9°C Without pronounced observable clinical manifestations Swelling / 2*2 cm / rash in the groin / body temperature: 38.6°C 1080 hours Rash Skin rash in the groin and armpit Without pronounced observable clinical manifestations Swelling 3*2 cm in the thigh / body temperature: 38.8°C Without pronounced observable clinical manifestations Swelling / 2*2 cm / rash in the groin / body temperature: 38.9°C 1104 hours Rash Skin rash in the groin and armpit / body temperature: 38.6°C Without pronounced observable clinical manifestations Swelling 2*2 cm in the thigh / body temperature: 38.9°C Without pronounced observable clinical manifestations Swelling reduction / rash reduction / body temperature: 39.1°C 1128 hours Rash reduction Skin rash in the groin and armpit Without pronounced observable clinical manifestations Swelling 2*2 cm in the thigh / body temperature: 38.8°C Without pronounced observable clinical manifestations Swelling reduction / rash reduction 1152 hours Rash reduction Skin rash in the groin and armpit Without pronounced observable clinical manifestations Swelling 2*2 cm in the thigh area Without pronounced observable clinical manifestations Swelling reduction / rash reduction 1176 hours Rash reduction Skin rash in the groin and armpit Without pronounced observable clinical manifestations Swelling reduction Without pronounced observable clinical manifestations Swelling reduction / rash reduction 1200 hours Rash reduction Skin rash in groin and armpit/body temperature: 38.7°C Without pronounced observable clinical manifestations Swelling reduction/body temperature: 38.8 Without pronounced observable clinical manifestations Swelling reduction/rash reduction/ body temperature: 38.9°C 1224 hours Rash reduction Skin rash in the groin and axilla / body temperature: 38.8 Without pronounced observable clinical manifestations Swelling reduction/body temperature: 39.0 Without pronounced observable clinical manifestations Swelling reduction / rash reduction / body temperature: 39.2 1248 hours Without pronounced observable clinical manifestations Skin rash in the groin and axilla / body temperature: 38.9 Without pronounced observable clinical manifestations Swelling reduction/body temperature: 38.9 Without pronounced observable clinical manifestations Swelling reduction / rash reduction / body temperature: 38.8 1272 hours Without pronounced observable clinical manifestations Skin rash in the groin and armpit Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 1296 hours Without pronounced observable clinical manifestations Skin rash in the groin and armpit Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 1512 hours Without pronounced observable clinical manifestations Skin rash in the groin and armpit Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 24 at 48 mg/kg (deep IM injection) are shown in Table 75.

Time point D1501 D1502 D1502 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 24 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.8 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 48 hours Without pronounced observable clinical manifestations Red spot next to the saphenous vein of the hind limb / body temperature: 38.6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 72 hours Without pronounced observable clinical manifestations Red spot next to the saphenous vein of the hind limb / body temperature: 38.8 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 96 hours Without pronounced observable clinical manifestations Red spot next to the saphenous vein of the hind limb / body temperature: 39.2 Without pronounced observable clinical manifestations Swelling throughout the leg, induration 11 * 11 cm in the groin / redness / body temperature: 38.3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 120 hours Without pronounced observable clinical manifestations Reduction of red spot next to the saphenous vein of the hind limb / body temperature: 39.2 Without pronounced observable clinical manifestations Swelling throughout the leg, induration 11 * 11 cm in the groin / redness / body temperature: 38.7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 144 hours Red spot in the groin Swelling 8 * 5 cm on the inner surface of the thigh with redness / swelling on the back of the lower leg / Reduction of the red spot near the saphenous vein of the hind limb / body temperature: 39.6 Without pronounced observable clinical manifestations Swelling throughout the leg, induration 11 * 11 cm in the groin / redness / body temperature: 38.3 Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 8 * 8 cm / body temperature: 39.5 168 hours Red spot in the groin Swelling 8 * 5 cm on the inner surface of the thigh with redness / swelling on the back of the lower leg / Reduction of the red spot near the saphenous vein of the hind limb / body temperature: 39.6 Without pronounced observable clinical manifestations Swelling throughout the leg, induration 11 * 11 cm in the groin / redness / body temperature: 38.4 Red spot in the groin Swelling on the inner thigh 8 * 8 cm / body temperature: 39.3 192 hours Red spot in the groin Swelling 8 * 5 cm on the inner surface of the thigh with redness / swelling on the back of the lower leg / body temperature: 39.0 Without pronounced observable clinical manifestations Swelling throughout the leg, induration 9 * 9 cm in the groin / redness / body temperature: 38.5 Red spot in the groin Swelling on the inner surface of the thigh 8 * 8 cm / body temperature: 38.0 216 hours Red spot in the groin Swelling 8 * 5 cm / body temperature: 39.0 Without pronounced observable clinical manifestations Swelling throughout the leg, induration 9 * 9 cm in the groin / redness / body temperature: 38.4 Red spot in the groin Swelling on the inner surface of the thigh 8 * 8 cm / body temperature: 38.8 240 hours Red spot in the groin Swelling 8 * 5 cm / body temperature: 39.0 Without pronounced observable clinical manifestations Swelling 9 * 9 cm on the back and inner thigh / redness / body temperature: 38.9 Red spot in the groin Swelling on the inner surface of the thigh 8 * 8 cm / body temperature: 39.2 264 hours Red spot in the groin Swelling 6 * 5 cm / body temperature: 38.9 Without pronounced observable clinical manifestations Swelling 9 * 9 cm on the back and inner thigh / redness / body temperature: 38.3 Red spot in the groin Swelling on the inner surface of the thigh 8 * 8 cm / body temperature: 38.9 288 hours Red spot in the groin Swelling 6 * 5 cm / body temperature: 38.9 Without pronounced observable clinical manifestations Swelling 9 * 9 cm on the back and inner thigh / redness / body temperature: 38.5 Reducing the red spot in the groin area Swelling on the inner surface of the thigh 8 * 8 cm / body temperature: 38.8 Day 13 Red spot in the groin Swelling 6*5 cm Without pronounced observable clinical manifestations Swelling 9 * 9 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 8 * 8 cm Day 14 Red spot in the groin Swelling 3*4 cm Without pronounced observable clinical manifestations Swelling 9 * 8 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 8 * 7 cm Day 15 Red spot in the groin Swelling 3*4 cm Without pronounced observable clinical manifestations Swelling 8 * 8 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 8 * 7 cm Day 16 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 6 * 6 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 6 * 6 cm Day 17 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 6 * 5 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 4 * 5 cm Day 18 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 6 * 5 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 4 * 5 cm Day 19 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 5 * 5 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 4 * 5 cm Day 20 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 5 * 5 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 4 * 5 cm Day 21 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 4 * 4 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 3 * 3 cm Day 22 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 4 * 4 cm on the back and inner thighs Without pronounced observable clinical manifestations Swelling on the inner surface of the thigh 2 * 2 cm Day 23 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 4 * 4 cm on the back and inner thighs Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 24 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 4 * 3 cm on the back and inner thighs Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 25 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 4 * 3 cm on the back and inner thighs Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Red spot in the groin Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 3 * 3 cm on the back and inner thighs Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 42 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 49 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 56 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 63 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 45 at 30 mg/kg are shown in Table 76.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 45 at 48 mg/kg are shown in Table 77.

Time point D1501 D1502 D1503 L-TA R-carrier L-TA R-carrier L-TA R-carrier Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 42 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 49 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 56 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 45 at 96 mg/kg are shown in Table 78.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Loose stools approximately 20 ml Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 7 at 24 mg/kg are shown in Table 79.

Time point D1001 D1002 L-carrier R-TA L-carrier R-TA Day 0 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 4 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 5 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 6 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 7 at 48 mg/kg are shown in Table 80.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 24 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 8 at 48 mg/kg are shown in Table 81.

Time point D1501 D1502 D1503 L-carrier (sesame oil +1% benzyl alcohol) R-TA L-carrier (cottonseed oil + 1% benzyl alcohol) R-TA L-carrier (sesame oil +1% benzyl alcohol) R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 24 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Swelling 6*6.5 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Swelling 5*3.5 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Swelling 5*3 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 37 Several areas of rash on the inside and outside of the left hind limb Several areas of rash on the outer side of the right hind limb, Swelling 5 * 3 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 40 Several areas of rash on the inside and outside of the left hind limb, more on the inside Several areas of rash on the inside and outside of the right hind limb, more on the inside Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 1 at 48 mg/kg are shown in Table 82.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 24 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 2 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 3 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 7 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 14 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 21 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 35 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 3 at 48 mg/kg are shown in Table 83.

Time point D1001 D1002 D1003 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 4*3.5 Without pronounced observable clinical manifestations Swelling 4*3 Day 1 Mottled mild redness of the skin Swelling / 4 * 4.5 cm / body temperature: 39.2 ° C / 2 * 2 cm red spot on the edge of the shaved area Without pronounced observable clinical manifestations Induration / 7 * 5 cm / marked swelling / body temperature: 39.1 ° C Mottled mild redness of the skin Induration / 8 * 5.5 cm / severe swelling / redness of the skin / higher temperature than at the injection site on the left side / body temperature: 39.6 ° C Day 2 Swelling / 5 * 6 cm / patchy slight redness of the skin Induration/ 8*11*1 cm/ 1*1 red spot near the injection site/ 3*3 red spot with yellow secretion around the edge of the shaved area/ body temperature: 38.9°C Swelling / 3*3 cm Induration / 7 * 7 * 1.5 cm / pronounced swelling / body temperature: 39.3 ° C Swelling / 3 * 3 cm / patchy slight redness of the skin Induration / 8 * 5.5 * 1 cm / severe swelling / redness of the skin / higher temperature than at the injection site on the left side / body temperature: 39.7 ° C Day 3 Swelling / 5 * 6 cm / patchy slight reddening of the skin Induration/ 8*11*1 cm/ 3*3 red spot with yellow secretion around the edge of the shaved area/ body temperature: 39.1°C Swelling / 3 * 3 cm / patchy slight redness of the skin Induration / 9 * 11 * 1.5 cm / severe swelling / body temperature: 39.3 ° C Swelling / 3 * 3 cm / patchy slight redness of the skin Induration / 7*11*1 cm / severe swelling / redness of the skin / higher temperature than at the injection site on the left side / body temperature: 39.2°C Day 4 Swelling / 5 * 6 cm / patchy slight redness of the skin Induration/ 10*11*1 cm/ 3*3 red spot with yellow secretion around the edge of the shaved area/ body temperature: 38.5°C Swelling / 3 * 4 cm / patchy slight reddening of the skin Induration / 9 * 11 * 1.5 cm / pronounced swelling / body temperature: 38.5 ° C Swelling / 3 * 3 cm / patchy slight redness of the skin Induration/ 8*12*1.5 cm/ Severe swelling/ reddening of the skin/ higher temperature than at the injection site on the left side/ another zone of induration at a distance of 4 cm from the injection site/ body temperature: 38.7°C Day 5 Swelling / 5 * 6 cm / patchy slight reddening of the skin Induration of the entire thigh / Swelling of the entire back of the leg / Soreness to touch, fluctuation, sensation of heat / 1.5 * 2 red spot with yellow secretion at the edge of the shaved area / body temperature: 38.6 ° C Swelling / 2.5 * 3.5 cm / patchy slight reddening of the skin Induration/ 9*10*2 cm/ Severe bloating/ Slight feeling of heat/ Body temperature: 38.7°C Swelling / 2 * 3.5 cm / patchy slight reddening of the skin Induration of the entire thigh / slight swelling of the entire back of the leg / another area of induration and severe swelling at a distance of 4 cm from the injection site / 3 * 3 cm red spot on the back of the thigh with slight swelling / body temperature: 39.0 ° C Day 6 Swelling / 5 * 6 cm / patchy slight redness of the skin / swelling of the entire back of the lower leg Induration of the entire thigh with slight redness / Swelling of the entire back of the leg / Soreness to touch, fluctuation, sensation of heat / 1.5 * 2 red spot with yellow secretion at the edge of the shaved area / body temperature: 38.4 ° C Swelling / 2 * 2 cm / patchy slight redness of the skin Induration/ 9*10*2 cm/ Severe bloating/ Slight feeling of heat/ Body temperature: 38.8°C Swelling / 2 * 3.5 cm / patchy slight reddening of the skin Induration of the entire thigh / slight swelling of the entire back of the leg / another area of induration and severe swelling at a distance of 4 cm from the injection site / 3 * 3 cm red spot on the back of the thigh with slight swelling / body temperature: 39.2 ° C Day 7 Swelling / 4 * 4 cm / patchy slight redness of the skin / swelling of the entire back of the lower leg Induration of the entire thigh with slight redness / swelling of the entire back of the lower leg / redness / sensation of heat / 1 * 1 red spot with a yellow secret along the edge of the shaved area / body temperature: 38.4 ° C Swelling / 2 * 2 cm / patchy slight redness of the skin Induration/ 9*10*2 cm/ Severe bloating/ Slight feeling of heat/ Body temperature: 38.6°C Swelling / 2 * 3.5 cm / patchy slight reddening of the skin Induration of the entire thigh / slight swelling of the entire back of the leg / another area of induration and severe swelling at a distance of 4 cm from the injection site / 3 * 3 cm red spot on the back of the thigh with slight swelling / body temperature: 39.5 ° C Day 8 Swelling / 4 * 4 cm / patchy slight redness of the skin / swelling of the entire back of the lower leg Induration of the entire thigh with slight redness / swelling of the entire back of the lower leg / redness / sensation of heat / 1 * 1 red spot with a yellow secretion at the edge of the shaved area / body temperature: 38.7 ° C Swelling / 2 * 2 cm / patchy slight redness of the skin Induration/ 9*10*2 cm/ Severe bloating/ Slight feeling of heat/ Body temperature: 38.7°C Swelling / 2 * 3.5 cm / patchy slight reddening of the skin Induration of the entire thigh / slight swelling of the entire back of the leg / another area of induration and severe swelling at a distance of 4 cm from the injection site / 3 * 3 cm red spot on the back of the thigh with slight swelling / body temperature: 39.4 °C Day 9 Swelling / 4 * 4 cm / patchy slight redness of the skin / swelling of the entire back of the lower leg Induration 11.5*9 cm in the thigh area with slight redness and a feeling of heat / slight swelling of the entire back of the lower leg / body temperature: 38.7 ° C Swelling reduction Induration / 9 * 10 * 2.5 cm / pronounced swelling / 4 scabs at 5 o'clock of the conventional dial at a distance of 2 cm from the induration zone and separation of one scab / slight redness / swelling of the entire back of the lower leg / body temperature: 38.3 °C Swelling / 2 * 3.5 cm / patchy slight reddening of the skin Induration/ 10*9*1.5 cm at the injection site/ slight swelling of the entire back of the lower leg/ another zone of induration and severe swelling at a distance of 4 cm from the injection site/ slight swelling in the thigh/ body temperature: 38.8°C Day 10 Swelling / 4 * 4 cm / patchy slight redness of the skin / swelling of the entire back of the lower leg Induration 11*9 cm in the thigh area with slight redness and a feeling of heat / slight swelling of the entire back of the lower leg / body temperature: 38.4°C Swelling reduction Induration / 9 * 10 * 2.5 cm / pronounced swelling / 4 scabs at 5 o'clock of the conventional dial at a distance of 2 cm from the induration zone and separation of two scabs / slight redness / swelling of the entire back of the lower leg / body temperature: 38.3 °C Swelling / 2 * 3.5 cm / patchy slight reddening of the skin Induration/ 10*9*1.5 cm at the injection site/ slight swelling of the entire back of the lower leg/ another zone of induration and severe swelling at a distance of 4 cm from the injection site/ slight swelling in the thigh/ body temperature: 38.9°C Day 11 Swelling / 4 * 4 cm / patchy slight redness of the skin / swelling of the entire back of the lower leg Induration 9*6 cm in the thigh area with slight redness and a feeling of heat / slight swelling of the entire back of the lower leg / body temperature: 38.8 ° C Swelling reduction Induration / 9 * 10 * cm / 3 yellow scabs at 5 o'clock of the conventional dial at a distance of 2 cm from the induration zone and separation of two scabs / slight redness / body temperature: 38.9 ° C Swelling Induration and sensation of heat/ 10*9*1.5 cm at the injection site/ another area of induration and pronounced swelling 4 cm from the injection site/ slight swelling in the thigh/ body temperature: 39.4°C Day 12 Swelling/ 4*4 cm Induration 9*6 cm in the thigh area with slight redness and a feeling of heat / body temperature: 38.0°C Swelling reduction Induration / 9*10 cm / 3 yellow scabs at 5 o'clock of the conventional dial at a distance of 2 cm from the induration zone and separation of two scabs / slight redness / body temperature: 38.5 ° C Swelling Induration and sensation of heat / 10 * 9 * 1.5 cm at the injection site / another zone of induration and severe swelling at a distance of 4 cm from the injection site / 3 * 3 cm red spot on the back of the thigh with slight swelling / body temperature : 39.3°C Day 13 Swelling/ 4*4 cm Induration 9*5 cm in the thigh area with slight redness and a feeling of heat / body temperature: 38.4°C Swelling reduction Induration / 9*10 cm / Slight redness / Feeling hot / Body temperature: 38.8°C Swelling Induration and sensation of heat/ 10*9 cm at the injection site/ another zone of induration and severe swelling at a distance of 4 cm from the injection site/ 3*3 cm red spot on the back of the thigh with slight swelling/ body temperature: 39.9 °C Day 14 Swelling/ 4*4 cm Induration 9*5 cm at the injection site Without pronounced observable clinical manifestations Induration / 8*8 cm / Slight redness / Feeling hot / Body temperature: 38.3°C Without pronounced observable clinical manifestations Induration and feeling of heat/ 8*9 cm at the injection site/ another area of induration and severe swelling 4 cm from the injection site/ slight swelling in the thigh/ body temperature: 39.6°C Day 15 Swelling/ 4*4 cm Reduction of induration 9*5 cm at the injection site / body temperature: 37.8°C Without pronounced observable clinical manifestations Induration / 8*8 cm / Slight redness / Feeling hot / Body temperature: 37.8°C Without pronounced observable clinical manifestations Induration and sensation of heat/ 8*9 cm at the injection site/ another area of induration and severe swelling 4 cm from the injection site/ slight swelling in the thigh/ body temperature: 39.4°C Day 16 Swelling/ 4*4 cm Reduction of induration 9 * 5 cm at the injection site Without pronounced observable clinical manifestations Induration / 8*8 cm / slight redness / feeling of heat Without pronounced observable clinical manifestations Induration and sensation of heat/ 8*9 cm at the injection site/ slight swelling of the entire back of the lower leg/ another area of induration and severe swelling at a distance of 4 cm from the injection site/ slight swelling in the thigh/ body temperature: 39.6°C Day 17 Swelling/ 4*4 cm Induration reduction 9*5 cm at injection site/ body temperature: 38.3°C Without pronounced observable clinical manifestations Induration / 8*8cm / body temperature: 38.7°C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.5°C Day 18 Swelling reduction Reduction of induration 9*5 cm at the injection site / body temperature: 38.4°C Without pronounced observable clinical manifestations Induration / 8*8cm / body temperature: 38.8°C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.5°C Day 19 Swelling reduction Induration reduction 9*5 cm at injection site/ body temperature: 38.3°C Without pronounced observable clinical manifestations Induration / 8*8cm / body temperature: 38.7°C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.4°C Day 20 Swelling reduction Induration reduction 9*5 cm at injection site/ body temperature: 37.9°C Without pronounced observable clinical manifestations Induration / 8 * 8 cm / body temperature: 38.6 ° C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.6°C Day 21 Swelling reduction Induration reduction 9*5 cm at injection site/ body temperature: 38.6°C Without pronounced observable clinical manifestations Induration reduction / 8*8 cm / sensation of heat / body temperature: 38.5°C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.3°C Day 22 Swelling reduction Reduction of induration 8*5 cm at the injection site / body temperature: 38.8°C Without pronounced observable clinical manifestations Induration reduction / 8*8 cm / sensation of heat / body temperature: 39.3°C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.9°C Day 23 Swelling reduction Reduction of induration 8*5 cm at the injection site / body temperature: 38.3°C Without pronounced observable clinical manifestations Induration reduction / 8*8 cm / sensation of heat / body temperature: 38.8°C Without pronounced observable clinical manifestations Induration and feeling of heat / 8*9 cm at the injection site / slight swelling in the thigh / body temperature: 39.6°C Day 24 Without pronounced observable clinical manifestations Swelling 6*3 cm at injection site/ body temperature: 38.5°C Without pronounced observable clinical manifestations Swelling reduction / 6*8 cm / sensation of heat / body temperature: 39.1°C Without pronounced observable clinical manifestations Swelling and feeling of heat / 6*6 cm at the injection site / Body temperature: 39.5°C Day 25 Without pronounced observable clinical manifestations Swelling 6*3 cm at injection site/ body temperature: 38.4°C Without pronounced observable clinical manifestations Swelling reduction / 6*8 cm / sensation of heat / body temperature: 38.6°C Without pronounced observable clinical manifestations Swelling and feeling of heat / 6*6 cm at the injection site / Body temperature: 39.8°C Day 26 Without pronounced observable clinical manifestations Swelling 5*3 cm at injection site/ body temperature: 38.2°C Without pronounced observable clinical manifestations Swelling reduction / 6*8cm / body temperature: 38.6°C Without pronounced observable clinical manifestations Swelling 6*6 cm at injection site/ body temperature: 39.3°C Day 27 Without pronounced observable clinical manifestations Swelling 5*3 cm at injection site/ body temperature: 38.4°C Without pronounced observable clinical manifestations Swelling reduction / 6*6cm / body temperature: 38.6°C Without pronounced observable clinical manifestations Swelling 5*6 cm at injection site/ body temperature: 38.9°C Day 28 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling reduction / 3*5 cm Without pronounced observable clinical manifestations Swelling 3*5 cm at the injection site Day 29 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling reduction / 3*5 cm Without pronounced observable clinical manifestations Swelling 3*3 cm at the injection site Day 30 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling reduction / 3*3 cm Without pronounced observable clinical manifestations Swelling 3*3 cm at the injection site Day 31 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling reduction / 3*3 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 32 Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling reduction / 3*3cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 4 at 48 mg/kg are shown in Table 84.

Time point D1501 D1502 D1503 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 1 Slight swelling Whole leg induration/hot sensation/limping Without pronounced observable clinical manifestations Swelling in the thigh area 6*7 cm Without pronounced observable clinical manifestations Whole leg induration Day 2 Slight swelling Induration of the whole leg / sensation of heat / lameness Without pronounced observable clinical manifestations Swelling in the thigh area 6 * 7 cm, induration on the inner surface of the thigh 7 * 7 cm / feeling of heat Without pronounced observable clinical manifestations Whole leg induration/feeling hot Day 3 Minor swelling Induration of the whole leg / sensation of heat / lameness Without pronounced observable clinical manifestations Swelling in the thigh area 6 * 7 cm, induration on the inner surface of the thigh 7 * 7 cm / feeling of heat Without pronounced observable clinical manifestations Whole leg induration/feeling hot Day 5 Minor swelling Thigh induration/feeling hot Without pronounced observable clinical manifestations Swelling in the thigh area 6 * 7 cm, induration on the inner surface of the thigh 7 * 7 cm / sensation of heat / eschar 1 * 0.5 cm to the right of the vulva Without pronounced observable clinical manifestations Whole leg induration/feeling hot Day 7 Slight swelling Thigh induration/feeling hot Without pronounced observable clinical manifestations Swelling in the thigh area 3 * 4 cm, induration on the inner surface of the thigh 7 * 7 cm / sensation of heat / eschar 1 * 0.5 cm to the right of the vulva Without pronounced observable clinical manifestations Whole leg induration/feeling hot Day 10 Without pronounced observable clinical manifestations Induration in the thigh area Without pronounced observable clinical manifestations Induration in the thigh / separation of the scab located near the vulva Without pronounced observable clinical manifestations Whole leg induration/feeling hot Day 12 Without pronounced observable clinical manifestations Induration in the thigh area Without pronounced observable clinical manifestations Induration in the thigh area 7 * 4 cm / separation of the scab located at the vulva Without pronounced observable clinical manifestations Induration in the thigh area 9*7 cm Day 17 Without pronounced observable clinical manifestations Induration in the thigh area Without pronounced observable clinical manifestations Induration in the thigh area 7*4 cm Without pronounced observable clinical manifestations Induration in the thigh area 9*6 cm Day 19 Without pronounced observable clinical manifestations Induration in the thigh area 8*5 cm Without pronounced observable clinical manifestations Induration in the thigh area 7*4 cm Without pronounced observable clinical manifestations Induration in the thigh area 7*4.5 cm Day 21 Without pronounced observable clinical manifestations Induration in the thigh area 8*4 cm Without pronounced observable clinical manifestations Induration in the thigh area 4*4 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 24 Without pronounced observable clinical manifestations Induration in the thigh area 7.5*4 cm Without pronounced observable clinical manifestations Induration in the thigh area 3.5*4 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 26 Without pronounced observable clinical manifestations Induration in the thigh area 6*4 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 28 Without pronounced observable clinical manifestations Induration in the thigh area 6*4 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 31 Without pronounced observable clinical manifestations Induration in the thigh area 6*4 cm Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Day 33 Without pronounced observable clinical manifestations Induration in the thigh area Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 5 at 48 mg/kg are shown in Table 85.

Time point D1001 D1002 L-carrier R-TA L-carrier R-TA 72 hours Without pronounced observable clinical manifestations Swelling 2*2.5cm Swelling 2*3cm Induration 6.5cm*4.5cm 96 hours Without pronounced observable clinical manifestations Swelling 3*3cm Swelling 4.5*2.5 cm Induration 6 cm*8 cm with slight redness 120 hours Without pronounced observable clinical manifestations Swelling 3*3cm Swelling 4.5*2.5 cm Induration 6 cm*8 cm with slight redness 144 hours Without pronounced observable clinical manifestations Swelling 3.5*3cm Swelling 4.5*2.5 cm Induration 6cm*8cm 168 hours Without pronounced observable clinical manifestations Induration 4*3 cm Swelling 4.5*2.5 cm Induration 6cm*8cm 192 hours Without pronounced observable clinical manifestations Induration 4*3 cm Swelling 4.5*2.5 cm Induration 6cm*8cm 216 hours Without pronounced observable clinical manifestations Induration 4*3 cm Swelling 4.5*2.5 cm Induration 6cm*8cm 240 hours Without pronounced observable clinical manifestations Swelling 2 * 3 cm / swelling with pus in the groin Swelling 3.5*2cm Induration 7cm*8cm 264 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.6 ° C Swelling 2*2cm Induration 7cm*8cm/ sensation of heat/body temperature: 39.5°C 288 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.6 ° C Swelling 2*2cm Induration 6cm*8cm/body temperature: 39.5°C 312 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.0 ° C Swelling 2*2cm Induration reduction / 6 cm * 8 cm / body temperature: 38.5 ° C 336 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.4 ° C Minor swelling 2*2 cm Induration reduction 3cm*4cm/body temperature: 38.7°C 360 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.1 ° C Without pronounced observable clinical manifestations Induration reduction 3cm*4cm/body temperature: 39.1°C 384 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.7 ° C Without pronounced observable clinical manifestations Induration reduction 3cm*4cm/body temperature: 39.3°C 408 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Induration reduction 3cm*4cm/body temperature: 39.4°C 432 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.0 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 38.9°C 456 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 38.9°C 480 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.0 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 39.2°C 504 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 38.7°C 528 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.2 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 37.9°C 552 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.4 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 39.2°C 576 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.4 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/ body temperature: 39.0°C 600 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.3 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 38.6°C 624 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.6 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 38.9°C 648 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.3 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 38.5°C 672 hours Without pronounced observable clinical manifestations Without severe observable clinical manifestations / body temperature: 38.3 ° C Without pronounced observable clinical manifestations Swelling 3cm*3cm/body temperature: 39.3°C 696 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 720 hours Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations

Clinical data for dogs treated with Compound 35 at 48 mg/kg are shown in Table 86. Time point D1501 D1502 D1502 L-carrier R-TA L-carrier R-TA L-carrier R-TA 8 ocloc'k Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations Without pronounced observable clinical manifestations 24 hours Swelling 4*4cm Red spots in the groin Swelling 3*4 cm Without pronounced observable clinical manifestations Slight swelling Slight swelling / red spot near the saphenous vein of the hind limb 48 hours Swelling 4*4cm Red spots in the groin Swelling 3*4 cm Without pronounced observable clinical manifestations Slight swelling Slight swelling / red spot near the saphenous vein of the hind limb 72 hours Swelling 4*4cm Swelling of the whole leg; significant swelling in the thigh area measuring 10 * 11 cm / red spots in the groin / body temperature: 39.1 ° C Swelling 3*4 cm Swelling of the whole leg; significant swelling on the inner surface of the thigh measuring 15 * 13 * 3 cm and redness / body temperature: 39.6 ° C Swelling 5*5 cm Swelling of the whole leg; significant swelling in the thigh area measuring 8 * 12 cm / redness on the inner surface of the thigh / body temperature: 39.3 ° C 96 hours Swelling 4*4cm Swelling of the whole leg; significant swelling in the thigh area measuring 10 * 11 cm / red spots in the groin / body temperature: 39.0 ° C Swelling 3*4 cm Swelling of the whole leg; significant swelling on the inner thigh measuring 15 * 13 * 3 cm and redness / body temperature: 38.9 ° C Swelling 5*5 cm Swelling of the whole leg; significant swelling in the thigh area measuring 8 * 12 cm / induration and redness on the inner surface of the thigh / body temperature: 39.3 ° C 120 hours Swelling 4*4cm Swelling of the whole leg; significant swelling in the thigh area measuring 10 * 11 cm / red spots in the groin / body temperature: 39.1 ° C Swelling 3*4 cm Swelling of the whole leg; significant swelling on the inner surface of the thigh measuring 15 * 13 * 3 cm and redness / body temperature: 39.6 ° C Swelling 5*5 cm Swelling of the whole leg; significant swelling in the thigh area measuring 8 * 12 cm / induration and redness on the inner surface of the thigh / red spot on the back of the lower leg / body temperature: 39.4 ° C 144 hours Swelling 4*4cm Significant swelling in the thigh area measuring 10 * 11 cm / 5 * 4 cm induration on the back of the thigh with redness / 9 * 7 cm induration on the inner surface of the thigh with redness / red spots in the groin / body temperature: 38.7 ° C Swelling 3*4 cm Induration of the entire leg; significant swelling on the inner thigh measuring 15 * 13 * 3 cm and redness / body temperature: 38.9 ° C Swelling 5*5 cm Swelling of the whole leg; significant swelling in the thigh area measuring 8 * 12 cm / induration and redness on the inner surface of the thigh / red spot on the back of the lower leg / body temperature: 39.0 ° C 168 hours Swelling 4*4cm Significant swelling in the thigh area measuring 10 * 11 cm / 5 * 4 cm induration on the back of the thigh with redness / 9 * 7 cm induration on the inner surface of the thigh with redness / red spots in the groin / body temperature: 38.9 ° C Swelling 3*4 cm Induration of the entire leg; significant swelling on the inner thigh measuring 15 * 13 * 3 cm and redness / body temperature: 39.9 ° C Swelling 5*5 cm Swelling of the whole leg; significant swelling in the thigh area measuring 8 * 12 cm / induration on the back of the thigh with redness / induration 8 * 6 cm on the inner surface of the thigh / red spot on the back of the leg / body temperature: 39.3 ° C 192 hours Swelling 4*4cm Significant swelling in the thigh area measuring 10 * 11 cm / 5 * 4 cm induration on the back of the thigh with redness / 9 * 7 cm induration on the inner surface of the thigh with redness / red spots in the groin / body temperature: 38.2 ° C Swelling 3*4 cm Induration of the entire leg; pronounced induration on the inner surface of the thigh measuring 15 * 13 cm / body temperature: 38.7 ° C Swelling 5 * 5 cm / red spots Significant swelling in the thigh area measuring 8 * 12 cm / induration on the back of the thigh with redness / induration of 8 * 6 cm on the inner surface of the thigh / red spot on the back of the leg / body temperature: 39.2 ° C 216 hours Swelling 4*4cm Significant swelling in the thigh area measuring 10 * 11 cm / 5 * 4 cm induration on the back of the thigh with redness / 9 * 7 cm induration on the inner surface of the thigh with redness / red spots in the groin / body temperature: 38.6 ° C Swelling 3*4 cm Pronounced induration on the inner surface of the thigh measuring 13 * 9 cm / body temperature: 39.2 ° C Swelling 4 * 4 cm / red spots Induration on the back and inside of the thigh measuring 7*12 cm / red spot on the back of the lower leg / body temperature: 39.2°C 240 hours Swelling reduction 5*4 cm induration on hamstring with redness/ 9*7 cm induration on inner thigh with redness/ red patches in groin/body temperature: 38.2°C Swelling 3*4 cm Pronounced induration on the inner surface of the thigh measuring 13 * 9 cm / body temperature: 38.6 ° C Swelling 4 * 4 cm / red spots Induration on the back and inside of the thigh measuring 7*12 cm / red spot on the back of the lower leg / body temperature: 38.9°C 264 hours Swelling reduction 3*4 cm induration on hamstring with redness/ 6*5 cm induration on inner thigh with redness/ red patches in groin/ body temperature: 38.0°C Swelling 3*3cm Pronounced induration on the inner surface of the thigh measuring 10 * 9 cm / body temperature: 38.4 ° C Swelling 2 * 2 cm / red spots Induration on the back and inner thighs 6*6 cm / body temperature: 38.7°C 288 hours Without pronounced observable clinical manifestations 3*4 cm induration on hamstring with redness/ 6*5 cm induration on inner thigh with redness/ red patches in groin/body temperature: 38.4°C Swelling 3*3cm Pronounced induration on the inner surface of the thigh measuring 9 * 9 cm / body temperature: 38.6 ° C red spots Induration on the back and inner thighs 6*6 cm / body temperature: 38.7°C Day 13 Without pronounced observable clinical manifestations Induration 3*4 cm on the back of the thigh with redness / red spots in the groin Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 7 * 7 cm Without pronounced observable clinical manifestations Induration on the back and inner surface of the thigh measuring 6 * 6 cm Day 14 Without pronounced observable clinical manifestations Induration 3*4 cm on the back of the thigh with redness / red spots in the groin Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 7 * 6 cm Without pronounced observable clinical manifestations Induration on the back and inner surface of the thigh measuring 6 * 6 cm Day 15 Without pronounced observable clinical manifestations Induration 3*4 cm on the back of the thigh with redness / red spots in the groin Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 7 * 7 cm Without pronounced observable clinical manifestations Induration on the back and inner surface of the thigh measuring 6 * 6 cm Day 16 Without pronounced observable clinical manifestations Induration 3*4 cm on the back of the thigh with redness / red spots in the groin Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 6 * 6 cm Without pronounced observable clinical manifestations Induration on the back and inner surface of the thigh measuring 6 * 6 cm Day 17 Without pronounced observable clinical manifestations Induration 3*2 cm on the back of the thigh with redness / red spots in the groin Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 6 * 6 cm Without pronounced observable clinical manifestations Induration on the back and inner thighs *6 cm Day 18 Without pronounced observable clinical manifestations Induration 3*2 cm on the back of the thigh with redness / red spots in the groin Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 5 * 5 cm Without pronounced observable clinical manifestations Induration on the back and inner thighs 4*3 cm Day 19 Without pronounced observable clinical manifestations Redness/red patches in the groin area Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 4 * 5 cm Without pronounced observable clinical manifestations Induration on the back and inner thighs 2*3 cm Day 20 Without pronounced observable clinical manifestations Redness/red patches in the groin area Without pronounced observable clinical manifestations Pronounced induration on the inner surface of the thigh measuring 4 * 4 cm Without pronounced observable clinical manifestations Without pronounced induration

Claims (18)

1. A compound, or a pharmaceutically acceptable salt thereof, having the structure shown below:

where X represents O or CH ₂ . 2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that the compound is

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that the compound is

4. Pharmaceutical composition for the treatment of opioid dependence, containing a compound according to any one of paragraphs. 1-3, or a pharmaceutically acceptable salt thereof, as an active ingredient, and at least one pharmaceutically acceptable excipient. 5. Pharmaceutical composition according to claim 4, characterized in that the pharmaceutically acceptable excipient is cottonseed oil. 6. Pharmaceutical composition according to claim 4, characterized in that the pharmaceutically acceptable excipient is sesame oil. 7. A method of treating opioid dependence in a patient in need of such treatment, comprising administering a pharmaceutical composition containing a compound according to any one of paragraphs. 1-3, or a pharmaceutically acceptable salt thereof, as an active ingredient, and a pharmaceutically acceptable excipient. 8. The method according to p. 7, characterized in that the pharmaceutical composition is administered by injection. 9. The method according to p. 8, characterized in that the pharmaceutical composition is administered by intramuscular injection. 10. The method according to claim 9, wherein the intramuscular injection is a depot injection. 11. The use of a pharmaceutical composition according to any one of paragraphs. 4-6 for the treatment of opioid dependence in a patient in need of such treatment. 12. Use according to claim 11, characterized in that the pharmaceutical composition is administered by injection. 13. Use according to claim 12, characterized in that the pharmaceutical composition is administered by intramuscular injection. 14. Use according to claim 13, characterized in that the intramuscular injection is a depot injection.

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