WO2016109360A1

WO2016109360A1 - Dihydroquinazoline inhibitors of viral terminase

Info

Publication number: WO2016109360A1
Application number: PCT/US2015/067542
Authority: WO
Inventors: Chengzhi Zhang; Justin CHAKMA
Original assignee: Auspex Pharmaceuticals, Inc.
Priority date: 2014-12-29
Filing date: 2015-12-27
Publication date: 2016-07-07
Also published as: AR103316A1; TW201629043A

Abstract

The present invention relates to new dihydroquinazoline modulators of viral infection, pharmaceutical compositions thereof, and methods of use thereof.

Description

DIHYDROQUINAZOLINE INHIBITORS OF VIRAL TERMINASE

[0001] This application claims the benefit of United States Provisional Application No. 62/097,443, filed December 31, 2014, the disclosure of which is hereby incorporated by reference as if written herein in its entirety.

[0002] Disclosed herein are new dihydroquinazoline compounds and compositions and their application as pharmaceuticals for the treatment of disorders. Methods of inhibition of viral terminase activity in a subject are also provided for the treatment of disorders such as human cytomegalovirus infection, human cytomegalovirus infection in stem cell recipients, human cytomegalovirus infection in bone marrow transplant patients, human

cytomegalovirus infection in kidney transplant patients, viral infection, herpes viridae infection, cytomegalovirus infection, human cytomegalovirus infection in AIDS patients, human cytomegalovirus infection in organ transplant patients, human cytomegalovirus pneumonitis, human cytomegalovirus encephalitis, gastrointestinal human cytomegalovirus infection, systemic human cytomegalovirus infection, human cytomegalovirus infection in neonates, human cytomegalovirus infection in infants, human cytomegalovirus infection in pregnant women, human cytomegalovirus infection in immune-suppressed patients, human cytomegalovirus infection in cancer, and human cytomegalovirus infection-mediated tumors.

[0003] Letermovir (AIC 246; CAS # 917389-32-3; (4S)-8-fluoro-3,4-dihydro-2-[4-(3- methoxyphenyl)-l-piperazinyl]-3-[2-methoxy-5-(trifluoromethyl)phenyl]-4-quinazolineacetic acid; (S)-2-(8-fluoro-3-(2-methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- methoxyphenyl)piperazin-l-yl)-3,4-dihydroquinazolin-4-yl)acetic acid) is a viral terminase inhibitor. Letermovir is currently under investigation for the treatment of human

cytomegalovirus infection, human cytomegalovirus infection in stem cell recipients, human cytomegalovirus infection in bone marrow transplant patients, and human cytomegalovirus infection in kidney transplant patients. Letermovir has also shown promise in treating viral infection, herpes viridae infection, cytomegalovirus infection, human cytomegalovirus infection in AIDS patients, human cytomegalovirus infection in organ transplant patients, human cytomegalovirus pneumonitis, human cytomegalovirus encephalitis, gastrointestinal human cytomegalovirus infection, systemic human cytomegalovirus infection, human cytomegalovirus infection in neonates, human cytomegalovirus infection in infants, human cytomegalovirus infection in pregnant women, human cytomegalovirus infection in immune- suppressed patients, human cytomegalovirus infection in cancer, and human cytomegalovirus infection-mediated tumors. WO 2013127971 ; WO 2013127970; WO 2013127968; WO 2006133822; WO 2004096778; US 20090221822; US 7,196,086; CA 2865049: U.S. Pat. Appl. No. 14/381,625; Stoelben et al, Transplant International, 2014, 27, 77-86; and Chemaly et al, New Eng. J. Me -1789.

Letermovir

[0004] Letermovir is likely subject to extensive CYP45o-mediated oxidative metabolism. These, as well as other metabolic transformations, occur in part through polymorphically- expressed enzymes, exacerbating interpatient variability. Additionally, some metabolites of letermovir derivatives may have undesirable side effects. In order to overcome its short half- life, the drug likely must be taken several times per day, which increases the probability of patient incompliance and discontinuance, and fluctuating plasma levels which may not offer ideal treatment.

Deuterium Kinetic Isotope Effect

[0005] In order to eliminate foreign substances such as therapeutic agents, the animal body expresses various enzymes, such as the cytochrome P450 enzymes (CYPs), esterases, proteases, reductases, dehydrogenases, and monoamine oxidases, to react with and convert these foreign substances to more polar intermediates or metabolites for renal excretion. Such metabolic reactions frequently involve the oxidation of a carbon-hydrogen (C-H) bond to either a carbon-oxygen (C-O) or a carbon-carbon (C-C) π-bond. The resultant metabolites may be stable or unstable under physiological conditions, and can have substantially different pharmacokinetic, pharmacodynamic, and acute and long-term toxicity profiles relative to the parent compounds. For most drugs, such oxidations are generally rapid and ultimately lead to administration of multiple or high daily doses.

[0006] The relationship between the activation energy and the rate of reaction may be quantified by the Arrhenius equation, k = Ae^{"Eact RT}. The Arrhenius equation states that, at a given temperature, the rate of a chemical reaction depends exponentially on the activation energy (E_act). [0007] The transition state in a reaction is a short lived state along the reaction pathway during which the original bonds have stretched to their limit. By definition, the activation energy E_act for a reaction is the energy required to reach the transition state of that reaction. Once the transition state is reached, the molecules can either revert to the original reactants, or form new bonds giving rise to reaction products. A catalyst facilitates a reaction process by lowering the activation energy leading to a transition state. Enzymes are examples of biological catalysts.

[0008] Carbon-hydrogen bond strength is directly proportional to the absolute value of the ground-state vibrational energy of the bond. This vibrational energy depends on the mass of the atoms that form the bond, and increases as the mass of one or both of the atoms making the bond increases. Since deuterium (D) has twice the mass of protium (¾), a C-D bond is stronger than the corresponding ( ¾ bond. If a C^H bond is broken during a rate- determining step in a chemical reaction (i.e. the step with the highest transition state energy), then substituting a deuterium for that protium will cause a decrease in the reaction rate. This phenomenon is known as the Deuterium Kinetic Isotope Effect (DKIE). The magnitude of the DKIE can be expressed as the ratio between the rates of a given reaction in which a C^H bond is broken, and the same reaction where deuterium is substituted for protium. The DKIE can range from about 1 (no isotope effect) to very large numbers, such as 50 or more.

Substitution of tritium for hydrogen results in yet a stronger bond than deuterium and gives numerically larger isotope effects

[0009] Deuterium (²H or D) is a stable and non-radioactive isotope of hydrogen which has approximately twice the mass of protium (¾), the most common isotope of hydrogen. Deuterium oxide (D2O or "heavy water") looks and tastes like H2O, but has different physical properties.

[0010] When pure D2O is given to rodents, it is readily absorbed. The quantity of deuterium required to induce toxicity is extremely high. When about 0-15% of the body water has been replaced by D2O, animals are healthy but are unable to gain weight as fast as the control (untreated) group. When about 15-20% of the body water has been replaced with D2O, the animals become excitable. When about 20-25% of the body water has been replaced with D2O, the animals become so excitable that they go into frequent convulsions when stimulated. Skin lesions, ulcers on the paws and muzzles, and necrosis of the tails appear. The animals also become very aggressive. When about 30% of the body water has been replaced with D2O, the animals refuse to eat and become comatose. Their body weight drops sharply and their metabolic rates drop far below normal, with death occurring at about 30 to about 35% replacement with D2O. The effects are reversible unless more than thirty percent of the previous body weight has been lost due to D2O. Studies have also shown that the use of D2O can delay the growth of cancer cells and enhance the cytotoxicity of certain antineoplastic agents.

[0011] Deuteration of pharmaceuticals to improve pharmacokinetics (PK),

pharmacodynamics (PD), and toxicity profiles has been demonstrated previously with some classes of drugs. For example, the DKIE was used to decrease the hepatotoxicity of halothane, presumably by limiting the production of reactive species such as trifluoroacetyl chloride. However, this method may not be applicable to all drug classes. For example, deuterium incorporation can lead to metabolic switching. Metabolic switching occurs when xenogens, sequestered by Phase I enzymes, bind transiently and re-bind in a variety of conformations prior to the chemical reaction (e.g., oxidation). Metabolic switching is enabled by the relatively vast size of binding pockets in many Phase I enzymes and the promiscuous nature of many metabolic reactions. Metabolic switching can lead to different proportions of known metabolites as well as altogether new metabolites. This new metabolic profile may impart more or less toxicity. Such pitfalls are non-obvious and are not predictable a priori for any drug class.

[0012] Letermovir is a viral terminase inhibitor. The carbon-hydrogen bonds of letermovir contain a naturally occurring distribution of hydrogen isotopes, namely ¾ or protium (about 99.9844%), ²H or deuterium (about 0.0156%), and ¾ or tritium (in the range between about 0.5 and 67 tritium atoms per 10¹⁸ protium atoms). Increased levels of deuterium incorporation may produce a detectable Deuterium Kinetic Isotope Effect (DKIE) that could effect the pharmacokinetic, pharmacologic and/or toxicologic profiles of such letermovir in comparison with the compound having naturally occurring levels of deuterium.

[0013] Based on discoveries made in our laboratory, as well as considering the literature, letermovir is likely metabolized in humans at the methoxy groups, the piperazine ring, the alpha and beta carbons to the carboxylic acid, and the dihydroquinazoline ring. The current approach has the potential to prevent metabolism at these sites. Other sites on the molecule may also undergo transformations leading to metabolites with as -yet-unknown

pharmacology/toxicology. Limiting the production of these metabolites has the potential to decrease the danger of the administration of such drugs and may even allow increased dosage and/or increased efficacy. All of these transformations can occur through polymorphically- expressed enzymes, exacerbating interpatient variability. Further, some disorders are best treated when the subject is medicated around the clock or for an extended period of time. For all of the foregoing reasons, a medicine with a longer half-life may result in greater efficacy and cost savings. Various deuteration patterns can be used to (a) reduce or eliminate unwanted metabolites, (b) increase the half-life of the parent drug, (c) decrease the number of doses needed to achieve a desired effect, (d) decrease the amount of a dose needed to achieve a desired effect, (e) increase the formation of active metabolites, if any are formed, (f) decrease the production of deleterious metabolites in specific tissues, and/or (g) create a more effective drug and/or a safer drug for polypharmacy, whether the polypharmacy be intentional or not. The deuteration approach has the strong potential to slow the metabolism of letermovir and attenuate interpatient variability.

[0014] Novel compounds and pharmaceutical compositions, certain of which have been found to inhibit viral terminase have been discovered, together with methods of synthesizing and using the compounds, including methods for the treatment of viral infection-mediated disorders in a patient by administering the compounds.

[0015] Accordingly, provided herein are compounds of structural Formula I:

or a salt thereof, wherein:

Ri and R2 are independently selected from the group consisting of -CH3, -CH2D, -

R3-R25 are independently selected from the group consisting of hydrogen and deuterium; and at least one of R1-R25 is deuterium or contains deuterium.

[0016] Also provided are enantiomers of compounds of Formula I, designated Formulas la and lb:

(lb).

In certain embodiments of the present invention, compounds have structural

(la)

or a salt thereof, wherein:

Ri and R2 are independently selected from the group consisting of -CH3, -CH2D,

R3-R25 are independently selected from the group consisting of hydrogen and deuterium; and

at least one of R1-R25 is deuterium or contains deuterium.

[0018] In certain embodiments, R9 is hydrogen.

[0019] In certain embodiments, Ri is -CD3.

[0020] In certain embodiments, R₂ is -CD3.

[0021] In certain embodiments, Ri and R2 are -CD3.

[0022] In certain embodiments, R3-R5 and R22-R25 are hydrogen.

[0023] In certain embodiments, R9. R3-R5, and R22-R25 are hydrogen.

[0024] In certain embodiments, R11-R13 are hydrogen.

[0025] In certain embodiments, R6 is hydrogen.

[0026] In certain embodiments, R7 and R8 are hydrogen.

[0027] In certain embodiments, R6-R8 are hydrogen.

[0028] In certain embodiments, four of R14-R21 are hydrogen.

[0029] In certain embodiments, six of R14-R21 are hydrogen.

[0030] In certain embodiments, R14-R21 are hydrogen.

[0031] In certain embodiments, R7, R8, and R14-R21 are hydrogen.

[0032] In certain embodiments, R6-R8 and R14-R21 are hydrogen. [0033] In certain embodiments, R.6 is deuterium.

[0034] In certain embodiments, Rj and Rs are deuterium.

[0035] In certain embodiments, R6-Rs are deuterium.

[0036] In certain embodiments, four of R14-R21 are deuterium.

[0037] In certain embodiments, six of R14-R21 are deuterium.

[0038] In certain embodiments, R14-R21 are deuterium.

[0039] In certain embodiments, R7, Rs, and R14-R21 are deuterium.

[0040] In certain embodiments, R6-Rs and R14-R21 are deuterium.

[0041] In certain embodiments, the compound is a prodrug of a compound of Formula I having one of formulas la, Ila, or Ilia below, and R9 is chosen from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, and hexyl.

[0042] Also provided herein are embodiments according to each of the embodiments above, wherein:

every other substituent among R3-R25 not specified as deuterium is hydrogen; and if either or both of Ri and R2 is not specified to be -CD3, then it is (they are) -CH3.

[0043] In certain embodiments compounds have structural Formula II:

(II)

or a salt thereof, wherein:

R6, R7, Rs, and R11-R21 are independently selected from the group consisting of hydrogen and deuterium; and

at least one of Ri, R2, R6, R7, Rs, and R11-R21 is deuterium or contains deuterium.

[0044] In certain embodiments, Ri is -CD3. [0045] In certain embodiments, R₂ is -CD₃.

[0046] In certain embodiments, Ri and R2 are -CD3.

[0047] In certain embodiments, R11-R13 are hydrogen.

[0048] In certain embodiments, R6 is hydrogen.

[0049] In certain embodiments, R7 and Rs are hydrogen.

[0050] In certain embodiments, R6-R8 are hydrogen.

[0051] In certain embodiments, four of R14-R21 are hydrogen.

[0052] In certain embodiments, six of R14-R21 are hydrogen.

[0053] In certain embodiments, R14-R21 are hydrogen.

[0054] In certain embodiments, R7, Rs, and R14-R21 are hydrogen.

[0055] In certain embodiments, R6-R8 and R14-R21 are hydrogen.

[0056] In certain embodiments, R6 is deuterium.

[0057] In certain embodiments, R7 and Rs are deuterium.

[0058] In certain embodiments, R6-R8 are deuterium.

[0059] In certain embodiments, four of R14-R21 are deuterium.

[0060] In certain embodiments, six of R14-R21 are deuterium.

[0061] In certain embodiments, R14-R21 are deuterium.

[0062] In certain embodiments, R7, Rs, and R14-R21 are deuterium.

[0063] In certain embodiments, R6-R8 and R14-R21 are deuterium.

[0064] In certain embodiments, the compound is a prodrug of a compound of Formula I having one of formulas la, Ila, or Ilia below, and R9 is chosen from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, and hexyl.

[0065] Also provided herein are embodiments according to each of the embodiments above, wherein:

every other substituent among R6, R7, Rs, and Rn-R2i not specified as deuterium is hydrogen; and

if either or both of Ri and R2 is not specified to be -CD3, then it is (they are) -CH3.

[0066] In certain embodiments, compounds have structural Formula III:

or a salt thereof, wherein:

Ri and R2 are independently selected from the group consisting of -CH3, -CH2D,

R6 and R14-R21 are independently selected from the group consisting of hydrogen deuterium; and

at least one of Ri, R2, R6, and R14-R21 is deuterium or contains deuterium.

[0067] In certain embodiments, Ri is -CD₃.

[0068] In certain embodiments, R2 is -CD₃.

[0069] In certain embodiments, Ri and R2 are -CD3.

[0070] In certain embodiments, R6 is hydrogen.

[0071] In certain embodiments, R7 and Rs are hydrogen.

[0072] In certain embodiments, R6-R8 are hydrogen.

[0073] In certain embodiments, four of R14-R21 are hydrogen.

[0074] In certain embodiments, six of R14-R21 are hydrogen.

[0075] In certain embodiments, R14-R21 are hydrogen.

[0076] In certain embodiments, R7, Rs, and R14-R21 are hydrogen.

[0077] In certain embodiments, R6-R8 and R14-R21 are hydrogen.

[0078] In certain embodiments, R6 is deuterium.

[0079] In certain embodiments, R7 and Rs are deuterium.

[0080] In certain embodiments, R6-R8 are deuterium.

[0081] In certain embodiments, four of R14-R21 are deuterium.

[0082] In certain embodiments, six of R14-R21 are deuterium.

[0083] In certain embodiments, R14-R21 are deuterium.

[0084] In certain embodiments, R7, Rs, and R14-R21 are deuterium. [0085] In certain embodiments, R.6-R.8 and R14-R21 are deuterium.

[0086] Also provided herein are embodiments according to each of the embodiments above, wherein:

every other substituent among R6 and R14-R21 not specified as deuterium is hydrogen; and

[0087] In certain embodiments of the present invention, compounds have structural Formula IV:

(IV)

or a salt thereof, wherein:

R2 is selected from the group consisting of -CH3, -CH2D, -CD2H, and -CD3;

at least one of R2-R25 is deuterium or contains deuterium.

[0088] In certain embodiments of the present invention, compounds have structural Formula V:

(V)

or a salt thereof, wherein:

Ri is selected from the group consisting of -CH3, -CH2D, -CD2H, and -CD3;

at least one of Ri and R3-R25 is deuterium or contains deuterium.

[0089] In certain embodiments of the present invention, compounds have structural Formula VI:

or a salt thereof, wherein: R3-R25 are independently selected from the group consisting of hydrogen and deuterium; and

at least one of R3-R25 is deuterium or contains deuterium.

[0090] Also provided is a compound chosen from the Examples and compounds disclosed herein.

[0091] In certain embodiments are provided compounds as disclosed herein, wherein at least one of R1-R25 independently has deuterium enrichment of no less than about 1%. In certain embodiments are provided compounds as disclosed herein, wherein at least one of Ri- R25 independently has deuterium enrichment of no less than about 10%. In certain embodiments are provided compounds as disclosed herein, wherein at least one of R1-R25 independently has deuterium enrichment of no less than about 50%. In certain embodiments are provided compounds as disclosed herein, wherein at least one of R1-R25 independently has deuterium enrichment of no less than about 90%. In certain embodiments are provided compounds as disclosed herein, wherein at least one of R1-R25 independently has deuterium enrichment of no less than about 95%. In certain embodiments are provided compounds as disclosed herein, wherein at least one of R1-R25 independently has deuterium enrichment of no less than about 98%.

[0092] The compounds as disclosed herein may also contain less prevalent isotopes for other elements, including, but not limited to, ¹ C or ¹⁴C for carbon, S, ⁴S, or ⁶S for sulfur, ¹⁵N for nitrogen, and ¹⁷0 or ¹⁸0 for oxygen.

[0093] In certain embodiments, the compound disclosed herein may expose a patient to a maximum of about 0.000005% D2O or about 0.00001% DHO, assuming that all of the C-D bonds in the compound as disclosed herein are metabolized and released as D2O or DHO. In certain embodiments, the levels of D2O shown to cause toxicity in animals is much greater than even the maximum limit of exposure caused by administration of the deuterium enriched compound as disclosed herein. Thus, in certain embodiments, the deuterium-enriched compound disclosed herein should not cause any additional toxicity due to the formation of D2O or DHO upon drug metabolism.

[0094] In certain embodiments are provided compounds as disclosed herein wherein each position represented as D has deuterium enrichment of no less than about 1%. In certain embodiments are provided compounds as disclosed herein wherein each position represented as D has deuterium enrichment of no less than about 10%. In certain embodiments are provided compounds as disclosed herein wherein each position represented as D has deuterium enrichment of no less than about 50%. In certain embodiments are provided compounds as disclosed herein wherein each position represented as D has deuterium enrichment of no less than about 90%. In certain embodiments are provided compounds as disclosed herein wherein each position represented as D has deuterium enrichment of no less than about 95%. In certain embodiments are provided compounds as disclosed herein wherein each position represented as D has deuterium enrichment of no less than about 98%.

[0095] In certain embodiments, the deuterated compounds disclosed herein maintain the beneficial aspects of the corresponding non-isotopically enriched molecules while substantially increasing the maximum tolerated dose, decreasing toxicity, increasing the half- life (T1/2), lowering the maximum plasma concentration (Cmax) of the minimum efficacious dose (MED), lowering the efficacious dose and thus decreasing the non-mechanism-related toxicity, and/or lowering the probability of drug-drug interactions.

[0096] Compounds disclosed herein possess useful viral terminase inhibiting activity, and may be used in the treatment or prophylaxis of a disorder in which viral infection play an active role. Thus, certain embodiments also provide pharmaceutical compositions comprising one or more compounds disclosed herein together with a pharmaceutically acceptable carrier, as well as methods of making and using the compounds and compositions. Certain embodiments provide methods for inhibiting viral terminase. Other embodiments provide methods for treating a viral infection-mediated disorder in a patient in need of such treatment, comprising administering to the patient a therapeutically effective amount of a compound or composition according to the present invention. Also provided is the use of certain compounds disclosed herein for use in the manufacture of a medicament for the prevention or treatment of a disorder ameliorated by the inhibition of viral terminase.

[0097] Also provided is a method of treatment of a viral infection-mediated disorder comprising the administration of a therapeutically effective amount of a compound as disclosed herein to a patient in need thereof.

[0098] In certain embodiments, the disorder is selected from the group consisting of human cytomegalovirus infection, human cytomegalovirus infection in stem cell recipients, human cytomegalovirus infection in bone marrow transplant patients, human

[0099] In certain embodiments, the method of treatment further comprises the administration of an additional therapeutic agent.

[00100] In certain embodiments, the additional therapeutic agent is an antiviral selected from the group consisting of ganciclovir, valganciclovir, foscarnet, cidofovir, and valaciclovir.

[00101] In certain embodiments, the method of treatment results in at least one effect selected from the group consisting of:

a) decreased inter-individual variation in plasma levels of the compound or a metabolite thereof as compared to the non-isotopically enriched compound; b) increased average plasma levels of the compound per dosage unit thereof as compared to the non-isotopically enriched compound;

c) decreased average plasma levels of at least one metabolite of the compound per dosage unit thereof as compared to the non-isotopically enriched compound;

d) increased average plasma levels of at least one metabolite of the compound per dosage unit thereof as compared to the non-isotopically enriched compound; and

e) an improved clinical effect during the treatment in the subject per dosage unit thereof as compared to the non-isotopically enriched compound.

[00102] In certain embodiments, the method of treatment further results in at least two effects selected from the group consisting of:

d) increased average plasma levels of at least one metabolite of the compound per dosage unit thereof as compared to the non-isotopically enriched compound; and e) an improved clinical effect during the treatment in the subject per dosage unit thereof as compared to the non-isotopically enriched compound.

[00103] In certain embodiments, the method effects a decreased metabolism of the compound per dosage unit thereof by at least one polymorphically-expressed cytochrome P450 isoform in the subject, as compared to the corresponding non-isotopically enriched compound.

[00104] In certain embodiments, the cytochrome P450 isoform is selected from the group consisting of CYP2C8, CYP2C9, CYP2C19, and CYP2D6.

[00105] In certain embodiments, the compound is characterized by decreased inhibition of at least one cytochrome P450 or monoamine oxidase isoform in the subject per dosage unit thereof as compared to the non-isotopically enriched compound.

[00106] In certain embodiments, the cytochrome P450 or monoamine oxidase isoform is selected from the group consisting of CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2, CYP2R1, CYP2S1, CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11, CYP4B1, CYP4F2, CYP4F3, CYP4F8, CYP4F11, CYP4F12, CYP4X1, CYP4Z1, CYP5A1, CYP7A1, CYP7B1, CYP8A1, CYP8B1, CYP11A1, CYP11B1, CYP11B2, CYP17, CYP19, CYP21, CYP24, CYP26A1, CYP26B1, CYP27A1, CYP27B1, CYP39, CYP46, CYP51, MAO A, and MAOB.

[00107] In certain embodiments, the method reduces a deleterious change in a diagnostic hepatobiliary function endpoint, as compared to the corresponding non-isotopically enriched compound.

[00108] In certain embodiments, the diagnostic hepatobiliary function endpoint is selected from the group consisting of alanine aminotransferase ("ALT"), serum glutamic-pyruvic transaminase ("SGPT"), aspartate aminotransferase ("AST," "SGOT"), ALT/AST ratios, serum aldolase, alkaline phosphatase ("ALP"), ammonia levels, bilirubin, gamma-glutamyl transpeptidase ("GGTP," "γ-GTP," "GGT"), leucine aminopeptidase ("LAP"), liver biopsy, liver ultrasonography, liver nuclear scan, 5 '-nucleotidase, and blood protein.

[00109] Also provided is a compound, or a salt thereof, as disclosed herein for use as a medicament.

[00110] Also provided is a compound, or a salt thereof, as disclosed herein for use in the manufacture of a medicament for the prevention or treatment of a viral infection-mediated disorder. [00111] All publications and references cited herein are expressly incorporated herein by reference in their entirety. However, with respect to any similar or identical terms found in both the incorporated publications or references and those explicitly put forth or defined in this document, then those terms definitions or meanings explicitly put forth in this document shall control in all respects.

[00112] As used herein, the terms below have the meanings indicated.

[00113] The singular forms "a," "an," and "the" may refer to plural articles unless specifically stated otherwise.

[00114] The term "about," as used herein, is intended to qualify the numerical values which it modifies, denoting such a value as variable within a margin of error. When no particular margin of error, such as a standard deviation to a mean value given in a chart or table of data, is recited, the term "about" should be understood to mean that range which would encompass the recited value and the range which would be included by rounding up or down to that figure as well, taking into account significant figures.

[00115] When ranges of values are disclosed, and the notation "from ni ... to m" or "m- m" is used, where m and are the numbers, then unless otherwise specified, this notation is intended to include the numbers themselves and the range between them. This range may be integral or continuous between and including the end values.

[00116] The term "deuterium enrichment" refers to the percentage of incorporation of deuterium at a given position in a molecule in the place of hydrogen. For example, deuterium enrichment of 1% at a given position means that 1% of molecules in a given sample contain deuterium at the specified position. Because the naturally occurring distribution of deuterium is about 0.0156%, deuterium enrichment at any position in a compound synthesized using non-enriched starting materials is about 0.0156%. The deuterium enrichment can be determined using conventional analytical methods known to one of ordinary skill in the art, including mass spectrometry and nuclear magnetic resonance spectroscopy.

[00117] The term "is/are deuterium," when used to describe a given position in a molecule such as Ri-R.25, or the symbol "D", when used to represent a given position in a drawing of a molecular structure, means that the specified position is enriched with deuterium above the naturally occurring distribution of deuterium. The same is true of the term "contains deuterium," which is often used to refer to methyl groups which may be mono-, di- or trideuterated (e.g., such groups may be -CH2D, -CD2H, and -CD3, wherein the each position denoted D is enriched with deuterium above the naturally occurring distribution of deuterium). In one embodiment deuterium enrichment is no less than about 1 %, in another no less than about 5%, in another no less than about 10%, in another no less than about 20%, in another no less than about 50%, in another no less than about 70%, in another no less than about 80%, in another no less than about 90%, or in another no less than about 98% of deuterium at the specified position.

[001 18] The term "isotopic enrichment" refers to the percentage of incorporation of a less prevalent isotope of an element at a given position in a molecule in the place of the more prevalent isotope of the element.

[001 19] The term "non-isotopically enriched" refers to a molecule in which the percentages of the various isotopes are substantially the same as the naturally occurring percentages.

[00120] Asymmetric centers exist in the compounds disclosed herein. These centers are designated by the symbols "R" or "S," depending on the configuration of substituents around the chiral carbon atom. It should be understood that the invention encompasses all stereochemical isomeric forms, including diastereomeric, enantiomeric, and epimeric forms, as well as d-isomers and 1 -isomers, and mixtures thereof. Individual stereoisomers of compounds can be prepared synthetically from commercially available starting materials which contain chiral centers or by preparation of mixtures of enantiomeric products followed by separation such as conversion to a mixture of diastereomers followed by separation or recrystallization, chromatographic techniques, direct separation of enantiomers on chiral chromatographic columns, or any other appropriate method known in the art. Starting compounds of particular stereochemistry are either commercially available or can be made and resolved by techniques known in the art. Additionally, the compounds disclosed herein may exist as geometric isomers. The present invention includes all cis, trans, syn, anti, entgegen (E), and zusammen (Z) isomers as well as the appropriate mixtures thereof.

Additionally, compounds may exist as tautomers; all tautomeric isomers are provided by this invention. Additionally, the compounds disclosed herein can exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. In general, the solvated forms are considered equivalent to the unsolvated forms.

[00121] The term "bond" refers to a covalent linkage between two atoms, or two moieties when the atoms joined by the bond are considered to be part of larger substructure. A bond may be single, double, or triple unless otherwise specified. A dashed line between two atoms in a drawing of a molecule indicates that an additional bond may be present or absent at that position. [00122] The term "disorder" as used herein is intended to be generally synonymous, and is used interchangeably with, the terms "disease" and "condition" (as in medical condition), in that all reflect an abnormal condition of the human or animal body or of one of its parts that impairs normal functioning, is typically manifested by distinguishing signs and symptoms.

[00123] The terms "treat," "treating," and "treatment" are meant to include alleviating or abrogating a disorder or one or more of the symptoms associated with a disorder; or alleviating or eradicating the cause(s) of the disorder itself. As used herein, reference to "treatment'Of a disorder is intended to include prevention. The terms "prevent,"

"preventing," and "prevention" refer to a method of delaying or precluding the onset of a disorder; and/or its attendant symptoms, barring a subject from acquiring a disorder or reducing a subject's risk of acquiring a disorder.

[00124] The term "therapeutically effective amount" refers to the amount of a compound that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disorder being treated. The term "therapeutically effective amount" also refers to the amount of a compound that is sufficient to elicit the biological or medical response of a cell, tissue, system, animal, or human that is being sought by a researcher, veterinarian, medical doctor, or clinician.

[00125] The term "subject" refers to an animal, including, but not limited to, a primate (e.g., human, monkey, chimpanzee, gorilla, and the like), rodents (e.g., rats, mice, gerbils, hamsters, ferrets, and the like), lagomorphs, swine (e.g., pig, miniature pig), equine, canine, feline, and the like. The terms "subject" and "patient" are used interchangeably herein in reference, for example, to a mammalian subject, such as a human patient.

[00126] The term "combination therapy" means the administration of two or more therapeutic agents to treat a therapeutic disorder described in the present disclosure. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule having a fixed ratio of active ingredients or in multiple, separate capsules for each active ingredient. In addition, such administration also encompasses use of each type of therapeutic agent in a sequential manner. In either case, the treatment regimen will provide beneficial effects of the drug combination in treating the disorders described herein.

[00127] The term "viral terminase" refers to a class of enzymes that play essential roles as components of molecular motors that package viral DNA into capsids. Herpesviral DNA packaging is a complex process involving binding and cleavage of DNA containing the specific DNA-packaging motifs, pacl and pac2, and packaging of the resulting unit-length genomes into preformed procapsids. This process is believed to be mediated by two packaging proteins, the terminase subunits. In the case of human cytomegalovirus the terminase consists of the proteins pUL56 and pUL89. While pUL56 (i) mediates the specific binding to pac sequences on the concatamers, (ii) provides energy for the translocation of the DNA to the procapsids and (iii) associates itself with the capsid for enabling the entry of the DNA into the procapsid, pUL89 is mainly required to effect DNA cleavage.

[00128] The term "viral infection-mediated disorder," refers to a disorder that is characterized by abnormal viral terminase activity or viral infection. A viral infection- mediated disorder may be completely or partially mediated by modulating viral terminase. In particular, a viral infection-mediated disorder is one in which inhibition of viral terminase results in some effect on the underlying disorder e.g., administration of a viral terminase inhibitor results in some improvement in at least some of the patients being treated.

[00129] The term "viral terminase inhibitor," refers to the ability of a compound disclosed herein to alter the function of viral terminase. An inhibitor may block or reduce the activity of viral terminase by forming a reversible or irreversible covalent bond between the inhibitor and viral terminase or through formation of a noncovalently bound complex. Such inhibition may be manifest only in particular cell types or may be contingent on a particular biological event. The term "inhibit" or "inhibition" also refers to altering the function of viral terminase by decreasing the probability that a complex forms between viral terminase and a natural substrate. In some embodiments, inhibition of reverse transcriptase may be assessed using the methods described in US 7, 196,086 and Lischka et al, Antimicrob. Agents Chemother., 2010, 54(3), 1290-1297.

[00130] The term "therapeutically acceptable" refers to those compounds (or salts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitable for use in contact with the tissues of patients without excessive toxicity, irritation, allergic response, immunogenecity, are commensurate with a reasonable benefit/risk ratio, and are effective for their intended use.

[00131] The term "pharmaceutically acceptable carrier," "pharmaceutically acceptable excipient," "physiologically acceptable carrier," or "physiologically acceptable excipient" refers to a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each component must be "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of a pharmaceutical formulation. It must also be suitable for use in contact with the tissue or organ of humans and animals without excessive toxicity, irritation, allergic response, immunogenecity, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

[00132] The terms "active ingredient," "active compound," and "active substance" refer to a compound, which is administered, alone or in combination with one or more

pharmaceutically acceptable excipients or carriers, to a subject for treating, preventing, or ameliorating one or more symptoms of a disorder.

[00133] The terms "drug," "therapeutic agent," and "chemotherapeutic agent" refer to a compound, or a pharmaceutical composition thereof, which is administered to a subject for treating, preventing, or ameliorating one or more symptoms of a disorder.

[00134] The term "release controlling excipient" refers to an excipient whose primary function is to modify the duration or place of release of the active substance from a dosage form as compared with a conventional immediate release dosage form.

[00135] The term "nonrelease controlling excipient" refers to an excipient whose primary function do not include modifying the duration or place of release of the active substance from a dosage form as compared with a conventional immediate release dosage form.

[00136] The term "prodrug" refers to a compound functional derivative of the compound as disclosed herein and is readily convertible into the parent compound in vivo. Prodrugs are often useful because, in some situations, they may be easier to administer than the parent compound. They may, for instance, be bioavailable by oral administration whereas the parent compound is not. The prodrug may also have enhanced solubility in pharmaceutical compositions over the parent compound. A prodrug may be converted into the parent drug by various mechanisms, including enzymatic processes and metabolic hydrolysis. Prodrugs may include esters of carboxylic acids, such as, for example, compounds of Formula la:

(la)

or a salt thereof, wherein:

R3- R8 and R10-R25 are independently selected from the group consisting of hydrogen and deuterium;

R9 is chosen from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, and hexyl; and

at least one of R1-R25 is deuterium or contains deuterium.

[00137] Prodrugs of compounds of Formulas Ila and Ilia,

(Ila) (Ilia) wherein R9 is chosen from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, and hexyl, and all other substituents are as described for formulas II and III, are also provided.

[00138] The compounds disclosed herein can exist as therapeutically acceptable salts. The term "therapeutically acceptable salt," as used herein, represents salts or zwitterionic forms of the compounds disclosed herein which are therapeutically acceptable as defined herein. The salts can be prepared during the final isolation and purification of the compounds or separately by reacting the appropriate compound with a suitable acid or base. Therapeutically acceptable salts include acid and basic addition salts.

[00139] Suitable acids for use in the preparation of pharmaceutically acceptable salts include, but are not limited to, acetic acid, 2,2-dichloroacetic acid, acylated amino acids, adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid, 4- acetamidobenzoic acid, boric acid, (+)-camphoric acid, camphorsulfonic acid, (+)-(l S)- camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamic acid, dodecylsulfuric acid, ethane- 1 ,2-disulfonic acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid, D-glucuronic acid, L-glutamic acid, a-oxo-glutaric acid, gly colic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, (+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid, maleic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-l,5-disulfonic acid, l -hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid, saccharic acid, salicylic acid, 4-amino- salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, and valeric acid.

[00140] Suitable bases for use in the preparation of pharmaceutically acceptable salts, including, but not limited to, inorganic bases, such as magnesium hydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, or sodium hydroxide; and organic bases, such as primary, secondary, tertiary, and quatemary, aliphatic and aromatic amines, including L-arginine, benethamine, benzathine, choline, deanol, diethanolamine, diethylamine, dimethylamine, dipropylamine, diisopropylamine, 2-(diethylamino)-ethanol, ethanolamine, ethylamine, ethylenediamine, isopropylamine, N-methyl-glucamine, hydrabamine, 1H- imidazole, L-lysine, morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine, piperazine, propylamine, pyrrolidine, l -(2-hydroxyethyl)-pyrrolidine, pyridine, quinuclidine, quinoline, isoquinoline, secondary amines, triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine, 2-amino-2-(hydroxymethyl)-l,3-propanediol, and tromethamine.

[00141] While it may be possible for the compounds of the subject invention to be administered as the raw chemical, it is also possible to present them as a pharmaceutical composition. Accordingly, provided herein are pharmaceutical compositions which comprise one or more of certain compounds disclosed herein, or one or more pharmaceutically acceptable salts, prodrugs, or solvates thereof, together with one or more pharmaceutically acceptable carriers thereof and optionally one or more other therapeutic ingredients. Proper formulation is dependent upon the route of administration chosen. Any of the well-known techniques, carriers, and excipients may be used as suitable and as understood in the art; e.g. , in Remington's Pharmaceutical Sciences. The pharmaceutical compositions disclosed herein may be manufactured in any manner known in the art, e.g. , by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or compression processes. The pharmaceutical compositions may also be formulated as a modified release dosage form, including delayed-, extended-, prolonged-, sustained-, pulsatile-, controlled-, accelerated- and fast-, targeted-, programmed-release, and gastric retention dosage forms. These dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art

[00142] The compositions include those suitable for oral, parenteral (including

subcutaneous, intradermal, intramuscular, intravenous, intraarticular, and intramedullary), intraperitoneal, transmucosal, transdermal, rectal and topical (including dermal, buccal, sublingual and intraocular) administration although the most suitable route may depend upon for example the condition and disorder of the recipient. The compositions may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. Typically, these methods include the step of bringing into association a compound of the subject invention or a pharmaceutically salt, prodrug, or solvate thereof ("active ingredient") with the carrier which constitutes one or more accessory ingredients. In general, the compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.

[00143] Formulations of the compounds disclosed herein suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient may also be presented as a bolus, electuary or paste.

[00144] Pharmaceutical preparations which can be used orally include tablets, push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Tablets may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with binders, inert diluents, or lubricating, surface active or dispersing agents. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be formulated so as to provide slow or controlled release of the active ingredient therein. All formulations for oral administration should be in dosages suitable for such administration. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.

[00145] The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in powder form or in a freeze- dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or sterile pyrogen-free water, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets of the kind previously described. [00146] Formulations for parenteral administration include aqueous and non-aqueous (oily) sterile injection solutions of the active compounds which may contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or

triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.

[00147] In addition to the formulations described previously, the compounds may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or

hydrophobic materials (for example as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.

[00148] For buccal or sublingual administration, the compositions may take the form of tablets, lozenges, pastilles, or gels formulated in conventional manner. Such compositions may comprise the active ingredient in a flavored basis such as sucrose and acacia or tragacanth.

[00149] The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g. , containing conventional suppository bases such as cocoa butter, polyethylene glycol, or other glycerides.

[00150] Certain compounds disclosed herein may be administered topically, that is by non- systemic administration. This includes the application of a compound disclosed herein externally to the epidermis or the buccal cavity and the instillation of such a compound into the ear, eye and nose, such that the compound does not significantly enter the blood stream. In contrast, systemic administration refers to oral, intravenous, intraperitoneal and intramuscular administration.

[00151] Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site of inflammation such as gels, liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose. [00152] For administration by inhalation, compounds may be delivered from an insufflator, nebulizer pressurized packs or other convenient means of delivering an aerosol spray. Pressurized packs may comprise a suitable propellant such as

dichlorodifiuoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. Alternatively, for administration by inhalation or insufflation, the compounds according to the invention may take the form of a dry powder composition, for example a powder mix of the compound and a suitable powder base such as lactose or starch. The powder composition may be presented in unit dosage form, in for example, capsules, cartridges, gelatin or blister packs from which the powder may be administered with the aid of an inhalator or insufflator.

[00153] Preferred unit dosage formulations are those containing an effective dose, as herein below recited, or an appropriate fraction thereof, of the active ingredient.

[00154] Compounds may be administered orally or via injection at a dose of from 0.1 to 500 mg/kg per day. The dose range for adult humans is generally from 5 mg to 2 g/day. Tablets or other forms of presentation provided in discrete units may conveniently contain an amount of one or more compounds which is effective at such dosage or as a multiple of the same, for instance, units containing 5 mg to 500 mg, usually around 10 mg to 200 mg.

[00155] The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration.

[00156] The compounds can be administered in various modes, e.g. orally, topically, or by injection. The precise amount of compound administered to a patient will be the

responsibility of the attendant physician. The specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diets, time of administration, route of administration, rate of excretion, drug combination, the precise disorder being treated, and the severity of the disorder being treated. Also, the route of administration may vary depending on the disorder and its severity.

[00157] In the case wherein the patient's condition does not improve, upon the doctor's discretion the administration of the compounds may be administered chronically, that is, for an extended period of time, including throughout the duration of the patient's life in order to ameliorate or otherwise control or limit the symptoms of the patient's disorder. [00158] In the case wherein the patient's status does improve, upon the doctor's discretion the administration of the compounds may be given continuously or temporarily suspended for a certain length of time (i.e., a "drug holiday").

[00159] Once improvement of the patient's conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, can be reduced, as a function of the symptoms, to a level at which the improved disorder is retained. Patients can, however, require intermittent treatment on a long-term basis upon any recurrence of symptoms.

[00160] Disclosed herein are methods of treating a viral infection-mediated disorder comprising administering to a subject having or suspected to have such a disorder, a therapeutically effective amount of a compound as disclosed herein or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[00161] Viral infection-mediated disorders, include, but are not limited to, human cytomegalovirus infection, human cytomegalovirus infection in stem cell recipients, human cytomegalovirus infection in bone marrow transplant patients, human cytomegalovirus infection in kidney transplant patients, viral infection, herpes viridae infection,

cytomegalovirus infection, human cytomegalovirus infection in AIDS patients, human cytomegalovirus infection in organ transplant patients, human cytomegalovirus pneumonitis, human cytomegalovirus encephalitis, gastrointestinal human cytomegalovirus infection, systemic human cytomegalovirus infection, human cytomegalovirus infection in neonates, human cytomegalovirus infection in infants, human cytomegalovirus infection in pregnant women, human cytomegalovirus infection in immune-suppressed patients, human cytomegalovirus infection in cancer, and human cytomegalovirus infection-mediated tumors, and/or any disorder which can lessened, alleviated, or prevented by administering a viral terminase inhibitor.

[00162] In certain embodiments, a method of treating a viral infection-mediated disorder comprises administering to the subject a therapeutically effective amount of a compound of as disclosed herein, or a pharmaceutically acceptable salt, solvate, or prodrug thereof, so as to affect: (1) decreased inter-individual variation in plasma levels of the compound or a metabolite thereof; (2) increased average plasma levels of the compound or decreased average plasma levels of at least one metabolite of the compound per dosage unit; (3) decreased inhibition of, and/or metabolism by at least one cytochrome P450 or monoamine oxidase isoform in the subject; (4) decreased metabolism via at least one polymorphically- expressed cytochrome P450 isoform in the subject; (5) at least one statistically-significantly improved disorder-control and/or disorder-eradication endpoint; (6) an improved clinical effect during the treatment of the disorder, (7) prevention of recurrence, or delay of decline or appearance, of abnormal alimentary or hepatic parameters as the primary clinical benefit, or (8) reduction or elimination of deleterious changes in any diagnostic hepatobiliary function endpoints, as compared to the corresponding non-isotopically enriched compound.

[00163] In certain embodiments, inter-individual variation in plasma levels of the compounds as disclosed herein, or metabolites thereof, is decreased; average plasma levels of the compound as disclosed herein are increased; average plasma levels of a metabolite of the compound as disclosed herein are decreased; inhibition of a cytochrome P450 or monoamine oxidase isoform by a compound as disclosed herein is decreased; or metabolism of the compound as disclosed herein by at least one polymorphically-expressed cytochrome P450 isoform is decreased; by greater than about 5%, greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, or by greater than about 50% as compared to the corresponding non-isotopically enriched compound.

[00164] Plasma levels of the compound as disclosed herein, or metabolites thereof, may be measured using the methods described in Stoelben et al, Transplant International, 2014, 27, 77-86, which is hereby incorporated by reference.

[00165] Examples of cytochrome P450 isoforms in a mammalian subject include, but are not limited to, CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2, CYP2R1, CYP2S1, CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11, CYP4B1, CYP4F2, CYP4F3, CYP4F8, CYP4F11, CYP4F12, CYP4X1, CYP4Z1, CYP5A1, CYP7A1, CYP7B1, CYP8A1, CYP8B1, CYP11A1, CYP11B1, CYP11B2, CYP17, CYP19, CYP21, CYP24, CYP26A1, CYP26B1, CYP27A1, CYP27B1, CYP39, CYP46, and CYP51.

[00166] Examples of monoamine oxidase isoforms in a mammalian subject include, but are not limited to, MAOA, and MAOB.

[00167] The inhibition of the cytochrome P450 isoform is measured by the method of Ko et al. {British Journal of Clinical Pharmacology, 2000, 49, 343-351). The inhibition of the MAOA isoform is measured by the method of Weyler et al. (J. Biol Chem. 1985, 260, 13199- 13207). The inhibition of the MAOB isoform is measured by the method of Uebelhack et al. (Pharmacopsychiatry, 1998, 31, 187-192).

[00168] Examples of polymorphically-expressed cytochrome P450 isoforms in a mammalian subject include, but are not limited to, CYP2C8, CYP2C9, CYP2C19, and CYP2D6. [00169] The metabolic activities of liver microsomes, cytochrome P450 isoforms, and monoamine oxidase isoforms are measured by the methods described herein.

[00170] Examples of improved disorder-control and/or disorder-eradication endpoints, or improved clinical effects include, but are not limited to, decrease in cytomegalovirus-DNA copy number, decreased viral load, and prophylaxis failure as determined by cytomegalovirus antigen and/or DNA detection. Stoelben et al, Transplant International, 2014, 27, 77-86; Chemaly et al, New Eng. J. Med., 2014, 370(90), 1781-1789.

[00171] Examples of diagnostic hepatobiliary function endpoints include, but are not limited to, alanine aminotransferase ("ALT"), serum glutamic-pyruvic transaminase

("SGPT"), aspartate aminotransferase ("AST" or "SGOT"), ALT/AST ratios, serum aldolase, alkaline phosphatase ("ALP"), ammonia levels, bilirubin, gamma-glutamyl transpeptidase ("GGTP," "γ-GTP," or "GGT"), leucine aminopeptidase ("LAP"), liver biopsy, liver ultrasonography, liver nuclear scan, 5 '-nucleotidase, and blood protein. Hepatobiliary endpoints are compared to the stated normal levels as given in "Diagnostic and Laboratory Test Reference", 4^th edition, Mosby, 1999. These assays are run by accredited laboratories according to standard protocol.

[00172] Besides being useful for human treatment, certain compounds and formulations disclosed herein may also be useful for veterinary treatment of companion animals, exotic animals and farm animals, including mammals, rodents, and the like. More preferred animals include horses, dogs, and cats.

Combination Therapy

[00173] The compounds disclosed herein may also be combined or used in combination with other agents useful in the treatment of viral infection-mediated disorders. Or, by way of example only, the therapeutic effectiveness of one of the compounds described herein may be enhanced by administration of an adjuvant (i.e., by itself the adjuvant may only have minimal therapeutic benefit, but in combination with another therapeutic agent, the overall therapeutic benefit to the patient is enhanced).

[00174] Such other agents, adjuvants, or drugs, may be administered, by a route and in an amount commonly used therefor, simultaneously or sequentially with a compound as disclosed herein. When a compound as disclosed herein is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound disclosed herein may be utilized, but is not required. [00175] In certain embodiments, the compounds disclosed herein can be combined with one or more antivirals selected from the group consisting of ganciclovir, valganciclovir, foscarnet, cidofovir, and valaciclovir.

[00176] The compounds disclosed herein can also be administered in combination with other classes of compounds, including, but not limited to, norepinephrine reuptake inhibitors (NRIs) such as atomoxetine; dopamine reuptake inhibitors (DARIs), such as

methylphenidate; serotonin-norepinephrine reuptake inhibitors (SNRIs), such as milnacipran; sedatives, such as diazepham; norepinephrine-dopamine reuptake inhibitor (NDRIs), such as bupropion; serotonin-norepinephrine-dopamine-reuptake-inhibitors (SNDRIs), such as venlafaxine; monoamine oxidase inhibitors, such as selegiline; hypothalamic phospholipids; endothelin converting enzyme (ECE) inhibitors, such as phosphoramidon; opioids, such as tramadol; thromboxane receptor antagonists, such as ifetroban; potassium channel openers; thrombin inhibitors, such as hirudin; hypothalamic phospholipids; growth factor inhibitors, such as modulators of PDGF activity; platelet activating factor (PAF) antagonists; antiplatelet agents, such as GPIIb/IIIa blockers (e.g., abdximab, eptifibatide, and tirofiban), P2Y(AC) antagonists (e.g., clopidogrel, ticlopidine and CS-747), and aspirin; anticoagulants, such as warfarin; low molecular weight heparins, such as enoxaparin; Factor Vila Inhibitors and Factor Xa Inhibitors; renin inhibitors; neutral endopeptidase (NEP) inhibitors;

vasopepsidase inhibitors (dual NEP -ACE inhibitors), such as omapatrilat and gemopatrilat; HMG CoA reductase inhibitors, such as pravastatin, lovastatin, atorvastatin, simvastatin, NK- 104 (a.k.a. itavastatin, nisvastatin, or nisbastatin), and ZD-4522 (also known as rosuvastatin, or atavastatin or visastatin); squalene synthetase inhibitors; fibrates; bile acid sequestrants, such as questran; niacin; anti-atherosclerotic agents, such as ACAT inhibitors; MTP

Inhibitors; calcium channel blockers, such as amlodipine besylate; potassium channel activators; alpha-muscarinic agents; beta-muscarinic agents, such as carvedilol and metoprolol; antiarrhythmic agents; diuretics, such as chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide, bendroflumethiazide, methylchlorothiazide,

trichioromethiazide, polythiazide, benzothlazide, ethacrynic acid, tricrynafen, chlorthalidone, furosenilde, musolimine, bumetanide, triamterene, amiloride, and spironolactone;

thrombolytic agents, such as tissue plasminogen activator (tPA), recombinant tPA, streptokinase, urokinase, prourokinase, and anisoylated plasminogen streptokinase activator complex (APSAC); anti-diabetic agents, such as biguanides (e.g. metformin), glucosidase inhibitors (e.g., acarbose), insulins, meglitinides (e.g., repaglinide), sulfonylureas (e.g., glimepiride, glyburide, and glipizide), thiozolidinediones (e.g. troglitazone, rosiglitazone and pioglitazone), and PPAR-gamma agonists; mineralocorticoid receptor antagonists, such as spironolactone and eplerenone; growth hormone secretagogues; aP2 inhibitors;

phosphodiesterase inhibitors, such as PDE III inhibitors (e.g., cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil, vardenafil); protein tyrosine kinase inhibitors;

antiinflammatories; antiproliferatives, such as methotrexate, FK506 (tacrolimus, Prograf), mycophenolate mofetil; chemotherapeutic agents; immunosuppressants; anticancer agents and cytotoxic agents (e.g., alkylating agents, such as nitrogen mustards, alkyl sulfonates, nitrosoureas, ethylenimines, and triazenes); antimetabolites, such as folate antagonists, purine analogues, and pyrridine analogues; antibiotics, such as anthracyclines, bleomycins, mitomycin, dactinomycin, and plicamycin; enzymes, such as L-asparaginase; farnesyl-protein transferase inhibitors; hormonal agents, such as glucocorticoids (e.g., cortisone), estrogens/antiestrogens, androgens/antiandrogens, progestins, and luteinizing hormone- releasing hormone anatagonists, and octreotide acetate; microtubule-disruptor agents, such as ecteinascidins; microtubule-stablizing agents, such as pacitaxel, docetaxel, and epothilones A-F; plant-derived products, such as vinca alkaloids, epipodophyllotoxins, and taxanes; and topoisomerase inhibitors; prenyl-protein transferase inhibitors; and cyclosporins; steroids, such as prednisone and dexamethasone; cytotoxic drugs, such as azathiprine and

cyclophosphamide; TNF-alpha inhibitors, such as tenidap; anti-TNF antibodies or soluble TNF receptor, such as etanercept, rapamycin, and leflunimide; and cyclooxygenase-2 (COX- 2) inhibitors, such as celecoxib and rofecoxib; and miscellaneous agents such as, hydroxyurea, procarbazine, mitotane, hexamethylmelamine, gold compounds, platinum coordination complexes, such as cisplatin, satraplatin, and carboplatin.

[00177] Thus, in another aspect, certain embodiments provide methods for treating viral infection- mediated disorders in a human or animal subject in need of such treatment comprising administering to the subject an amount of a compound disclosed herein effective to reduce or prevent the disorder in the subject, in combination with at least one additional agent for the treatment of the disorder that is known in the art. In a related aspect, certain embodiments provide therapeutic compositions comprising at least one compound disclosed herein in combination with one or more additional agents for the treatment of viral infection- mediated disorders.

General Synthetic Methods for Preparing Compounds

[00178] Isotopic hydrogen can be introduced into a compound as disclosed herein by synthetic techniques that employ deuterated reagents, whereby incorporation rates are pre- determined; and/or by exchange techniques, wherein incorporation rates are determined by equilibrium conditions, and may be highly variable depending on the reaction conditions. Synthetic techniques, where tritium or deuterium is directly and specifically inserted by tritiated or deuterated reagents of known isotopic content, may yield high tritium or deuterium abundance, but can be limited by the chemistry required. Exchange techniques, on the other hand, may yield lower tritium or deuterium incorporation, often with the isotope being distributed over many sites on the molecule.

[00179] The compounds as disclosed herein can be prepared by methods known to one of skill in the art and routine modifications thereof, and/or following procedures similar to those described in the Example section herein and routine modifications thereof, and/or procedures found in WO 2013127971 ; WO 2013127970; WO 2013127968; WO 2006133822; WO 2004096778; US 20090221822; US 7,196,086; CA 2865049: and U.S. Pat. Appl. No.

14/381,625, which are hereby incorporated in their entirety, and references cited therein and routine modifications thereof. Compounds as disclosed herein can also be prepared as shown in any of the following schemes and routine modifications thereof.

[00180] The following schemes can be used to practice the present invention. Any position shown as hydrogen may optionally be replaced with deuterium.

Scheme I



[00181] Compound 1 is treated with an appropriate phosgene equivalent, such as trichloromethyl chloroformate, in the presence of an appropriate base, such as 1,8- bis(dimethylamino)napthalene, in an appropriate solvent, such as dichloromethane, to give compound 2. Compound 2 is reacted with compound 3, in an appropriate solvent, such as acetonitrile, to give compound 4. Compound 4 is reacted with compound 5 in the presence of an appropriate catalyst, such as palladium acetate, in the presence of an appropriate acid catalyst, such as fuming sulfuric acid, in an appropriate solvent, such as acetic acid, under a 4% O2 / 96% N2 atmosphere, to give compound 6. Compound 6 is reacted with an appropriate base, such as l,8-diazabicyclo[5.4.0]undec-7-ene, in an appropriate solvent, such as acetone, at an elevated temperature, to give compound 7. Compound 9 is reacted with compound 10 in the presence of an appropriate catalyst, such as a combination of tris(dibenzylideneacetone)dipalladium and (2,2'-bis(diphenylphosphino)-l,l'-binaphthyl, in an appropriate solvent, such as toluene, at elevated temperature, to give compound 11.

Compound 7 is treated with an appropriate chlorinating agent, such as phosphorous oxy chloride, in the presence of an appropriate base, such as l,8-Diazabicyclo[5.4.0]undec-7- ene, in an appropriate solvent, such as chlorobenzene, at an elevated temperature, to give an intermediate 8, which is further reacted with compound 11 in the presence of an appropriate base, such as l,8-diazabicyclo[5.4.0]undec-7-ene, in an appropriate solvent, such as 1,4- dioxane, at an elevated temperature compound 12. Compound 12 is reacted with an appropriate chiral acid, such as (2S,3S)-2,3-bis[(4-methylbenzoyl)oxy]-succinic acid, in an appropriate solvent, such as ethyl acetate, to give compound 13. Compound 13 may be optionally recrystallized by dissolution in ethyl acetate at an elevated temperature followed by filtration and cooling. Compound 13 is treated with an appropriate base, such as sodium bicarbonate, in an appropriate solvent, such as a combination of methyl tert-butyl ether and water, to give compound 13 as the free base, which is further reacted with an appropriate base, such as sodium hydroxide, in an appropriate solvent, such as water, to give a compound of formula I. Alternatively, Compound 12 is reacted with an appropriate base, such as sodium hydroxide, in an appropriate solvent, such as dioxane and water, to give compound of formula I as a racemic mixture followed separation by chiral chromatographic techniques to provide compound of formula I optically pure.

[00182] Deuterium can be incorporated to different positions synthetically, according to the synthetic procedures as shown in Scheme I, by using appropriate deuterated

intermediates. For example, to introduce deuterium at R2-R5, compound 1 with the corresponding deuterium substitutions can be used. To introduce deuterium at one or more positions of R11-R13, compound 3 with the corresponding deuterium substitutions can be used. To introduce deuterium at one or more positions of R6-R7, compound 5 can be used. To introduce deuterium at Rs, acetic acid and/or sulfuric acid with the corresponding deuterium substitutions can be used. To introduce deuterium at one or more positions of R14-R21, compound 9 with the corresponding deuterium substitutions can be used. To introduce deuterium at one or more positions of Ri and R22-R25, compound 10 with the corresponding deuterium substitutions can be used.

[00183] Deuterium can be incorporated to various positions having an exchangeable proton, such as the carboxyl O-H, via proton-deuterium equilibrium exchange. For example, to introduce deuterium at R9 this proton may be replaced with deuterium selectively or non- selectively through a proton-deuterium exchange method known in the art.

[00184] The invention is further illustrated by the following examples.

EXAMPLE 1

Step 1 : Synthesis of 3-(2-bromo-6-fluorophenyl)-l-[2-methoxy-5- (trifluoromethy l)pheny 1] urea:

1 2

[00185] To a solution of triphosgene (10.5 g, 0.35 equiv) in dichloromethane (200.0 mL) was added TEA (108.0 g, 1.07 mol, 10.00 equiv) dropwise with stirring at 0 °C. To this was added a solution of 2-bromo-6-fluoroaniline (1, 20.0 g, 105.26 mmol, 1.00 equiv) in dichloromethane (100 mL) dropwise with stirring at 0 °C and then a solution of 2-methoxy-5- (trifluoromethyl)aniline (22.8 g, 119.28 mmol, 1.00 equiv) in dichloromethane (200 mL) was added dropwise with stirring at room-temperature. The resulting solution was stirred for 3 h at 25 °C under an inert atmosphere of nitrogen. The reaction progress was monitored by LCMS. The resulting mixture was then concentrated under vacuum and the residue was dissolved in 200 mL of ethyl acetate and then was washed with 100 mL of sodium bicarbonate and then was dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 20.1 g of 3-(2-bromo-6-fluorophenyl)-l-[2-methoxy-5-(trifluoromethyl)phenyl]urea (2) as a white solid. LC-MS: m/z = 407.25 [M+l]⁺ ¾ NMR (300 MHz, DMSO-de) δ: 8.89-8.82 (m, 2 H), 8.49-8.48 (m, 1 H), 7.55-7.20 (m, 5 H), 4.01 (s, 3 H).

Step 2: Methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2-oxo-l,2,3,4- tetrahydroquinazolin-4-yl]acetate:

[00186] To a solution of 3-(2-bromo-6-fluorophenyl)-l-[2-methoxy-5- (trifluoromethyl)phenyl]urea (2, 2.0 g, 4.91 mmol, 1.00 equiv) in CfbCN (10.0 mL), were added Pd(MeCN)₂Cl₂ (50.0 mg, 0.19 mmol, 0.04 equiv), P(o-Tol)₃ (120.0 mg, 0.39 mmol, 0.08 equiv), methyl prop-2-enoate (1.2 g, 13.94 mmol, 3.00 equiv), TEA (1.0 g, 9.88 mmol, 2.00 equiv). The final reaction mixture was irradiated with microwave radiation for 2 h at 110 °C. The reaction progress was monitored by LCMS. The solids were filtered out immediately at > 80 °C. The filtrate was cooled to room temperature. The solids were collected by filtration to afford 1.1 g of methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2- oxo-l,2,3,4-tetrahydroquinazolin-4-yl]acetate (3) as a white solid. LC-MS: m/z =413.10 [M+l]⁺. ¾ NMR (300 MHz, DMSO-de) δ: 9.78 (s , 1 H), 7.98-6.92 (m, 6 H), 5.17-5.13 (m, 1 H), 3.85 (s, 3 H), 3.43(s, 3H), 2.84-2.50 ( m , 2 H).

Step 3: Methyl 2-[2-chloro-8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-3,4-

[00187] To a solution of phosphoroyl trichloride (10 mL) was added methyl 2-[8-fluoro-3- [2-methoxy-5-(trifluoromethyl)phenyl]-2-oxo-l,2,3,4-tetrahydroquinazolin-4-yl]acetate (3, 2 g, 4.85 mmol, 1.00 equiv) in several batches. Then DIEA (1.3 g, 10.06 mmol, 2.00 equiv) was added drop wise with stirring. The resulting solution was stirred for 16 h at 120 °C under an inert atmosphere of nitrogen. The reaction progress was monitored by LCMS. The resulting mixture was then concentrated under vacuum. The residue was dissolved in 20 mL of ethyl acetate and washed with 1 x 20 mL of sodium bicarbonate, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 1.7 g of methyl 2-[2-chloro- 8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-3,4-dihydroquinazolin-4-yl]acetate

(Intermediate 1) as brown oil. LC-MS: m/z = 431.1 [M+l]⁺

Step 4: Methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2-[4-(3- methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4-yl]acetate

[00188] To a solution of Intermediate 1(1.7 g, 3.95 mmol, 1.00 equiv) in dioxane (7 mL) were added l-(3-methoxyphenyl)piperazine (1.2 g, 6.24 mmol, 1.50 equiv), DBU (1.2 g, 7.88 mmol, 2.00 equiv). The reaction mixture was irradiated with microwave radiation for 3 h at 130 °C. The reaction progress was monitored by LCMS. The resulting solution was diluted with 20 mL of water, extracted with 2 x 20 mL of ethyl acetate. The organic layers were combined and washed with 1 x 20 mL of HC1, dried over anhydrous sodium sulfate and concentrated under vacuum to afford 800 mg of methyl 2-[8-fluoro-3-[2-methoxy-5- (trifluoromethyl)phenyl]-2-[4-(3-methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4- yl] acetate (5) as yellow oil.

[00189] Step 5: (S)-2-(8-fluoro-3-(2-methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- methoxyphenyl)piperazin-l-yl)-3,4-dihydroquinazolin-4-yl)acetic acid

[00190] To a solution of methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2- [4-(3-methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4-yl]acetate (5, 600 mg) in dioxane (5 mL ) was added 1 mol/L NaOH (2.5 mL) and the mixture was stirred for 1 overnight at room-temperature and was monitored by LCMS. The reaction mixture was adjusted pH to 3 with 1 mol/L HC1 and extracted with 3 x 5 mL of ethyl acetate. The organic layers were combined and dried over anhydrous sodium, filtered and concentrated under vacuum. The crude product was purified by Prep-SFC with the following conditions:

Column, Chiralpak OD-H, 20*250mm, 20um; mobile phase, CO₂(70%), methanol(30%); Detector, UV 254 nm to afford 62.2 mg of the product as a light yellow solid. ¾ NMR (300 MHz, CDCh) δ: 7.69 - 6.31 (m , 10 H), 4.93 - 4.88 (m, 1 H), 3.74 (s, 6 H), 3.66 - 2.62 (m, 10 H). LC-MS: m/z = 573.15 [M+l]⁺.

EXAMPLE 2

(S)-2-(8-fluoro-3-(2-methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- (²H3)methoxyphenyl) azolin-4-yl)acetic acid

Intermediate 2: l-[3-(2H3)methoxyphenyl]piperazine

Intermediate 2-Step 1 : l-bromo-3-(²H3)methoxybenzene

[00191] To a solution of 3-bromophenol (5 g, 28.90 mmol, 1.00 equiv) in CH₃CN (100 mL) were added CD3I (5.1 g, 35.17 mmol, 1.20 equiv), potassium carbonate (8.0 g, 57.97 mmol, 2.00 equiv). The resulting solution was stirred for 5 h at 100 °C. The reaction mixture was cooled to toom temperature. The resulting mixture was concentrated under vacuum. The residue was dissolved in dichloromethane (100 mL) and washed with H2O (3 x 30 mL). The organic layer was collected and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 5 g of l-bromo-3-(²H3) methoxybenzene as yellow liquid.

Intermediate 2-Step 2: l-[3-(²H3)methoxyphenyl]piperazine

[00192] To a solution of l-bromo-3-(²H3) methoxybenzene (6.7 g, 35.25 mmol, 1.00 equiv) in tolene (100 mL) were added piperazine (6.1 g, 70.82 mmol, 2.00 equiv), Pd2(dba)3 (1.8 g, 1.74 mmol, 0.05 equiv), P(t-Bu)3 (2.05 g, 7.07 mmol, 0.20 equiv), NaOtBu (5.1 g, 53.12 mmol, 1.50 equiv), under a N2 atmosphere. The resulting solution was stirred for 15 hr at lOOoC in an oil bath. The reaction progress was monitored by LCMS. The solids were filtered out. The resulting mixture was concentrated under vacuum. The pH value of the solution was adjusted to 3 with HC1 (1 mol/L). The resulting solution was extracted with ethyl acetate (3x30 mL) and the aqueous layers combined, sodium hydroxide (1 mol/L) was employed to adjust the pH to 10. The resulting solution was extracted with ethyl acetate (3x30 mL) and the organic layers combined and dried over anhydrous sodium sulfate. This resulted in 5 g (73%) of l-[3-(²H3)methoxyphenyl]piperazine as brown oil.

Step 1 : Methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2-[4-(3- 2H3)methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4-yl]acetate

[00193] To a solution of Intermediate 1 (1.8 g, 4.18 mmol, 1.00 equiv) in dioxane (15 mL) were added l-(3-(²H3)methoxyphenyl)piperazine (Intermediate 2, 1.47 g, 7.53 mmol, 1.80 equiv), DBU (1.15 g, 7.53 mmol, 1.80 equiv). The reaction mixture was irradiated with microwave radiation for 3 h at 130 °C. The reaction progress was monitored by LCMS. The resulting solution was diluted with 20 mL of water and extracted with 2 x 20 mL of ethyl acetate. The organic layers were combined and washed with 1 x 20 mL of HC1, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 1.2 g of methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2-[4-(3-(²H3)methoxyphenyl)piperazin- l-yl]-3,4-dihydroquinazolin-4-yl]acetate as a brown solid. LC-MS: m/z = 590.20 [M+l]⁺

Step 2: (S)-2-(8-fluoro-3-(2-methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- 2H3)methoxyphenyl)piperazin-l-yl)- -dihydroquinazolin-4-yl)acetic acid

[00194] To a solution of methyl 2-[8-fluoro-3-[2-methoxy-5-(trifluoromethyl)phenyl]-2- [4-(3-(²H3)methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4-yl]acetate (1.1 g) in dioxane (30 mL ) was added 1 mol/L NaOH (4.0 ml ). The mixture was stirred for 1 overnight at room-temperature and was monitored by LCMS. The reaction mixture was then adjusted pH to 3 with 1 mol/L HC1, extracted with 3 x 10 mL of ethyl acetate. The organic layers were combined and dried over anhydrous sodium, filtered and concentrated under vacuum. The crude product was purified by Prep-SFC with the following conditions:

Column, Chiralpak OD-H, 20*250mm, 20 urn; mobile phase, CO₂(70%), methanol(30%); Detector, UV 254 nm to afford 192.9 mg of the product as a white solid. ¾ NMR (300 MHz, CDCh) δ: 9.80 (s , 1 H) ,7.70 - 6.30 (m , 10 H), 4.96 - 4.92 (m, 1 H), 3.74 (s, 3 H), 3.53 - 2.51 (m, 10 H). LC-MS: m/z = 576.20 [M+l]⁺.

EXAMPLE 3

(S)-2-(8-fluoro-3-(2-(²H₃)methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- methoxyphenyl)piperazin-l-yl)-3,4-dihydroquinazolin-4-yl)acetic acid

Preparation of Intermediate 3; 2-(²H3)methoxy-5-(trifluoromethyl)aniline

[00195] To a solution of CD3OD (40 mL) was added Na (2.2 g, 95.65 mmol, 2.00 equiv) in portions in 20 min. Then l-fluoro-2-nitro-4-(trifluoromethyl) benzene (10 g, 47.82 mmol, 1.00 equiv) was added in portions at 15 °C. The resulting solution was stirred for 3 h at 20 °C Then Palladium carbon (40 mg, 10%) was added to the mixture. The resulting solution was stirred for 15 h at 20 °C under a H2 atmosphere (2-4 atm). The reaction progress was monitored by LCMS. The solids were filtered out. The residue was dissolved in

dichloromethane (100 mL) and washed with H2O (2 x 40 mL). The organic layer was collected and dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to afford 7 g (75%) of 2-(²]¾) methoxy-5-(trifluoromethyl)aniline as a white solid. -(2-bromo-6-fluorophenyl)-l-[2-( ²H3 )methoxy-5-(trifluoromethyl)phenyl]urea

[00196] To a solution of triphosgene (1.25 g, 0.35 equiv) in dichloromethane (55.0 mL) was added TEA (14.5 mL 10.00 equiv) dropwise with stirring at 0 ⁰ C. Then a solution of 2- bromo-6-fiuoroaniline (1, 2.0 g, 10.53 mmol, 1.00 equiv) in dichloromethane (10 mL) was added dropwise with stirring at 0 °C. The reaction was allowed to warm to room-temperature and a solution of 2-(²H3)-methoxy-5-(trifluoromethyl)aniline (Intermediate 3, 2.04 g, 10.51 mmol, 1.00 equiv) in dichloromethane (20 mL) was added dropwise with stirring. The resulting solution was stirred for 3 h at 25 °C with an inert atmosphere of nitrogen. The reaction progress was monitored by LCMS. The resulting mixture was concentrated under vacuum and the residue was dissolved in 20 mL of ethyl acetate, washed with 10 mL of sodium bicarbonate, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 2.8 g of 3-(2-bromo-6-fluorophenyl)-l-[2-(²H3)methoxy-5- (trifluoromethyl)phenyl]urea (2) as a white solid. LC-MS: m/z = 410.05 [M+l]⁺ ¾ NMR (300 MHz, DMSO-de) δ: 8.91-8.81 (m, 2 H), 8.50-8.49 (m, 1 H), 7.56-7.20 (m, 5 H) .

Step 2: Methyl 2-[8-fluoro-3-[2-(²H₃)-methoxy-5-(trifluoromethyl)phenyl]-2-oxo-l,2,3,4- tetrahydroquinazolin-4-yl]acetate

[00197] To a solution of 3-(2-bromo-6-fluorophenyl)-l-[2-(²H₃)methoxy-5- (trifluoromethyl)phenyl]urea (2, 2.0 g, 4.88 mmol, 1.00 equiv) in CH3CN (10.0 mL), were added Pd(MeCN)₂Cl₂ (50.5 mg, 0.19 mmol, 0.04 equiv), P(o-Tol)₃ (119.0 mg, 0.39 mmol, 0.08 equiv), methyl prop-2-enoate (1.26 g, 14.64 mmol, 3.00 equiv), TEA (985 mg, 9.88 mmol, 2.00 equiv). The reaction mixture was irradiated with microwave radiation for 2 h at 110 °C. The reaction progress was monitored by LCMS. The solids were filtered out immediately at > 80 °C. The filtrate was cooled to room temperature. The solids were collected by filtration to afford 1.5 g of methyl 2-[8-fluoro-3-[2-(²H3)methoxy-5- (trifluoromethyl)phenyl]-2-oxo-l,2,3,4-tetrahydroquinazolin-4-yl]acetate (3) as a white solid. LC-MS: m/z =416.20 [M+l]⁺. ¾ NMR (300 MHz, DMSO-de) δ: 9.73 (s , 1 H), 7.73-6.93 (m, 6 H), 5.17-5.13 (m, 1 H), 3.41(s, 3H), 2.85-2.51 (m, 2 H).

Step 3: Methyl 2-[2-chloro-8-fluoro-3-[2-(²H3)methoxy-5-(trifluoromethyl)phenyl]-3,4- dihy droquinazolin-4-yl] acetate

3 Intermediate 4

[00198] To a solution of phosphoroyl trichloride (5.0 mL) was added methyl 2-[8-fluoro- 3-[2-(²H3)methoxy-5-(trifluoromethyl)phenyl]-2-oxo-l,2,3,4-tetrahydroquinazolin-4- yljacetate (3, 1.0 g, 2.41 mmol, 1.00 equiv) in several batches. Then DIEA (684 mg, 5.30 mmol, 2.20 equiv) was added dropwise with stirring. The resulting solution was stirred for 16 h at 120 °C with an inert atmosphere of nitrogen. The reaction progress was monitored by LCMS. The resulting mixture was concentrated under vacuum and the residue was dissolved in 20 mL of ethyl acetate, washed with 1 x 20 mL of sodium bicarbonate, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 700 mg of methyl 2-[2-chloro-8-fluoro-3-[2-(²H3)methoxy-5-(trifluoromethyl)phenyl]-3,4- dihydroquinazolin-4-yl] acetate (Intermediate 4) as brown oil. LC-MS: m/z = 434.1 [M+l]⁺

Step 4: Methyl 2-[8-fluoro-3-[2-(²H₃)methoxy-5-(trifluoromethyl)phenyl]-2-[4-(3- methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4-yl]acetate

[00199] To a solution of Intermediate 4 (1.0 g, 2.31 mmol, 1.00 equiv) in dioxane (4 mL) were added l-(3-methoxyphenyl)piperazine (800 mg, 4.16 mmol, 1.80 equiv), DBU (636 mg, 4.16 mmol, 1.80 equiv). The reaction mixture was irradiated with microwave radiation for 3 h at 130 °C. The reaction progress was monitored by LCMS. The resulting solution was diluted with 20 mL of water, extracted with 2 x 20 mL of ethyl acetate. The organic layers were combined, washed with 1 x 20 mL of HC1 (10%), dried over anhydrous sodium sulfate and concentrated under vacuum to afford 600 mg of methyl 2-[8-fluoro-3-[2-(²H3)methoxy-5- (trifluoromethy l)pheny 1] -2- [4-(3-m^

yl]acetate (5) as a brown solid. LC-MS: m/z = 590.30 [M+l]⁺

Step 5: (S)-2-(8-fluoro-3-(2-(²H₃)methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- methoxyphenyl)piperazin- 1 -yl)-3,4-dihy droquinazolin-4-yl)acetic acid

[00200] To a solution of methyl 2-[8-fluoro-3-[2-(²H₃)methoxy-5- (trifluoromethyl)phenyl]-2-[4-(3-methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4- yljacetate (5, 2.0 g) in dioxane (30 mL ) were added 1 mol/L NaOH (6.0 ml ), the mixture was stirred for 1 overnight at room-temperature and was monitored by LCMS. The reaction mixture was then adjusted pH to 3 with 1 mol/L HCl and then was extracted with 3 x 10 mL of ethyl acetate and the organic layers combined and dried over anhydrous sodium, filtered and concentrated under vacuum. The crude product was purified by Prep-SFC with the following conditions: Column, Chiralpak OD-H, 20*250mm, 20um; mobile phase, CC (70%), methanol(30%); Detector, UV 254 nm to afford 129 mg of the product as a light yellow solid. ¾ NMR (300 MHz, CDCh) δ: 7.70 - 6.30 (m , 10 H), 4.94 - 4.90 (m, 1 H), 3.77 (s, 3 H), 3.53 - 2.58 (m, 10 H). LC-MS: m/z = 576.15 [M+l]⁺.

EXAMPLE 4

(S)-2-(8-fluoro-3-(2-(²H₃)methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- (²H3)methoxyphenyl)piperazin-l-yl)-3,4-dihydroquinazolin-4-yl)acetic acid

-letermovir)

Step 1: Methyl 2-[8-fluoro-3-[2-(²H₃)methoxy-5-(trifluoromethyl)phenyl]-2-[4-(3- (²H3)methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin-4-yl]acetate

[00201] To a solution of methyl 2-[2-chloro-8-fluoro-3-[2-(²H₃)methoxy-5- (trifluoromethyl)phenyl]-3,4-dihydroquinazolin-4-yl]acetate (2, 2.2 g, 5.07 mmol, 1.00 equiv) in dioxane (15 mL) were added Intermediate 2 (1.78 g, 9.12 mmol, 1.80 equiv), DBU (1.4 g, 9.12 mmol, 1.80 equiv). The reaction mixture was irradiated with microwave radiation for 3 h at 130 °C. The reaction progress was monitored by LCMS. The resulting solution was diluted with 20 mL of water, extracted with 3 x 30 mL of ethyl acetate. The combined organic layers was washed with 2 x 20 mL of HC1, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford 1.2 g of methyl 2-[8-fiuoro-3-[2-(²H3) methoxy-5- (trifluoromethyl)phenyl]-2-[4-(3-(²H3)methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin- 4-yl]acetate (3) as yellow solid. LC-MS: m/z = 593.20 [M+l]⁺

Step 2: (S)-2-(8-fluoro-3-(2-(²H₃)methoxy-5-(trifluoromethyl)phenyl)-2-(4-(3- 2H3)methoxyphenyl)piperazin-l-yl)- -dih dro uinazolin-4- l acetic acid

[00202] To a solution of methyl 2-[8-fluoro-3-[2-(²H₃)methoxy-5-

(trifluoromethyl)phenyl]-2-[4-(3(²H3)-methoxyphenyl)piperazin-l-yl]-3,4-dihydroquinazolin- 4-yl]acetate (1.5 g) in dioxane (30 mL ) , was add 1 mol/L NaOH (4.5 ml ), the mixture was stirred for 1 overnight at room-temperature and was monitored by LCMS. The reaction mixture was then adjusted pH to 3 with 1 mol / L HC1 and then was extracted with 3 x 5 mL of ethyl acetate and the organic layers combined and dried over anhydrous sodium, filtered and concentrated under vacuum. The crude product was purified by Prep-SFC with the following conditions: Column, Chiralpak OD-H, 20*250mm, 20um; mobile phase,

CO₂(70%), methanol(30%); Detector, UV 254 nm to afford 100.9 mg of the product as a white solid. ¾ NMR (300 MHz, CDCh) δ: 9.35 (s , 1 H) ,7.97 - 6.28 (m , 10 H), 5.01 - 4.97 (m, 1 H), 3.55 - 2.48 (m, 10 H). LC-MS: m/z = 579.20 [M+l]⁺.

[00203] The following compounds can generally be made using the methods described above. All IUPAC names were generated using CambridgeSoft's ChemDraw 10.0. It is expected that these compounds when made will have activity similar to those described in the examples above.

49

50

51

52

53

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61

62

63

64 [00204] Changes in the metabolic properties of the compounds disclosed herein as compared to their non-isotopically enriched analogs can be shown using the following assays. Compounds listed above which have not yet been made and/or tested are predicted to have changed metabolic properties as shown by one or more of these assays as well.

Biological Activity Assays

In vitro Liver Microsomal Stability Assay

[00205] Human liver microsomal stability assays are conducted at 1.0 mg per mL liver microsome protein with NADPH (2mM, pH 7.4). Test compounds are typically prepared as solutions in acetonitrile with 5% DMSO and added to the assay mixture (0.5 uM, final concentration in incubation) to be incubated at 37 °C. Reactions are initiated with the addition of NADPH cofactor and are stopped at 0, 30, 60, 90 or 120 min after cofactor addition with stop reagent, acetonitrile. After quenching, plates containing samples are vibrated for 10 min (600 rpm/min) and then centrifuged at 5594 g for 15 min. Supernatant fractions are analyzed by LC-MS/MS to determine the percent remaining and estimate the degradation half-life of the test compounds. Results are given below in Table 1.

Table 1.

[00206] Liver microsomal stability assays may also be conducted at 1 mg per mL liver microsome protein with an NADPH-generating system in 2% NaHCC (2.2 mM NADPH, 25.6 mM glucose 6-phosphate, 6 units per mL glucose 6-phosphate dehydrogenase and 3.3 mM MgCh). Test compounds are prepared as solutions in 20% acetonitrile-water and added to the assay mixture (final assay concentration 5 microgram per mL) and incubated at 37 °C. Final concentration of acetonitrile in the assay should be <\%. Aliquots (50μί) are taken out at times 0, 15, 30, 45, and 60 min, and diluted with ice cold acetonitrile (200 μί) to stop the reactions. Samples are centrifuged at 12,000 RPM for 10 min to precipitate proteins. Supernatants are transferred to microcentrifuge tubes and stored for LC/MS/MS analysis of the degradation half-life of the test compounds.

In vitro metabolism using human cytochrome P450 enzymes

[00207] The cytochrome P450 enzymes are expressed from the corresponding human cDNA using a baculovirus expression system (e.g., from BD Biosciences, San Jose, CA). A 0.25 milliliter reaction mixture containing 0.8 milligrams per milliliter protein, 1.3 millimolar NADP⁺, 3.3 millimolar glucose-6-phosphate, 0.4 U/mL glucose-6-phosphate dehydrogenase, 3.3 millimolar magnesium chloride and 0.2 millimolar of a compound of Formula I, the corresponding non-isotopically enriched compound or standard or control in 100 millimolar potassium phosphate (pH 7.4) is incubated at 37 °C for 20 min. After incubation, the reaction is stopped by the addition of an appropriate solvent (e.g., acetonitrile, 20% trichloroacetic acid, 94% acetonitrile/6% glacial acetic acid, 70% perchloric acid, 94% acetonitrile/6% glacial acetic acid) and centrifuged (10,000 g) for 3 min. The supernatant is analyzed by HPLC/MS/MS. Compounds disclosed herein are expected to have activity in this assay as demonstrated by reduced metabolism by one or more cytochrome P450 enzymes of deuterated compound as compared to the non-isotopically enriched compound.

Table 2.

In vitro CYP3A4 Stability Assay

[00208] Recombinant CYP3A4 stability assays were conducted at an enzyme

concentration of 20 pmol per mL with NADPH (2mM, pH 7.4). Test compounds were prepared as acetonitrile with 5% DMSO and added to the assay mixture (luM, final concentration in incubation) to be incubated at 37 °C. Reactions were initiated with the addition of NADPH cofactor and stopped at 0, 30, 60, 90 or 120 min after cofactor addition with stop reagent, acetonitrile. After quenching, plates containing samples were vibrated for 10 min (600 rpm/min) and then centrifuged at 5594 g for 15 min. Supernatant fractions were analyzed by LC-MS/MS to determine the percent remaining and estimate the degradation half-life of the test compounds. Results are given below in Table 3.

Table 3.

In vitro Human Liver Microsomal Metabolite ID Assay

[00209] In vitro metabolite identification was conducted after incubating compounds (parent compound, final concentration 0.5 μΜ) with human liver microsomes at 37°C in 100 mM potassium phosphate buffer containing 1 mM EDTA in the presence of NADPH or without NADPH (w/o). Samples taken at 0 min and 120 min were quenched by using acetonitrile and analyzed using Xevo G2-S QTof system with positive-ion electrospray ionization. LC-UV-MS extract ion chromatograms (EIC) of the T omin and Tominwere compared to identify the major putative metabolites. The MS/MS spectra of compounds were also obtained to facilitate metabolite structure elucidation. Results are given below in Table 4. A reduced prevalence of the demethylation metabolite was noted and this is consistent with the increased stability with Examples 2 and 4. A complementary increase in the hydroxylation metabolite was also observed, indicating that the deuterated compounds' metabolism is quantitatively shifting from demethylation to hydroxylation.

Table 4.

Example 1 UV* 100.00% 85.73% + +

(letermovir) MS peak area 1.62E+04 1.53E+04 2.00E+02 4.84E+01

UV* 100.00% 86.09% + -

Example 2

MS peak area 2.04E+04 2.07E+04 6.74E+02 -

UV* 100.00% 100.00% + +

Example 3

MS peak area 1.05E+04 1.41E+04 3.50E+02 5.69E+01

UV* 100.00% 100.00% + -

Example 4

MS peak area 1.77E+04 2.10E+04 4.32E+02 -

Oral rat pharmacokinetics

[00210] Male rats (weight approximately 225g) were orally dosed with 50 mg/kg SD- 1086, SD-1087, SD-1088 and SD-1090 in 0.5% methylcellulose (10 mL/kg) in water after an overnight fast (n=3). Blood samples were obtained predose(O) and 0.083, 0.25, 0.5, 1, 2, 4, 8 and 24 hr post dose on K2EDTA and processed to plasma with sample handling procedures at on ice or at 4C. Plasma samples were stored at -70C for subsequent bioanalytical assay. Plasma samples were diluted as needed with plasma blanks (analyte free) as needed to stay with the range of calibration, then mixed with ACN which containing dexamethasone as an internal standard IS for protein precipitation. The mixture was vortexed for 5 min and centrifuged at 5800 rpm for 10 min. The supernatant was injected into LC-MS/MS with MRM detection to determine concentration of orally dosed compound. Plasma

concentrations were determined with a calibration curve. Plasma concentration profiles of 3 rats per group were amenable for pharmacokinetic parameter analysis with the exception of one rat.

Table 5.

Monoamine Oxidase A Inhibition and Oxidative Turnover [00211] The procedure is carried out using the methods described by Weyler, Journal of Biological Chemistry 1985, 260, 13199-13207, which is hereby incorporated by reference in its entirety. Monoamine oxidase A activity is measured spectrophotometrically by monitoring the increase in absorbance at 314 nm on oxidation of kynuramine with formation of 4-hydroxyquinoline. The measurements are carried out, at 30 °C, in 50mM NaPi buffer, pH 7.2, containing 0.2% Triton X-100 (monoamine oxidase assay buffer), plus 1 mM kynuramine, and the desired amount of enzyme in 1 mL total volume.

Monooamine Oxidase B Inhibition and Oxidative Turnover

[00212] The procedure is carried out as described in Uebelhack, Pharmacopsychiatry 1998, 37(5), 187-192, which is hereby incorporated by reference in its entirety.

Anti-Human Cytomegalovirus Cvtopathogenicitv Tests

[00213] The procedure is carried out as described in US 7,196,086, which is hereby incorporated by reference in its entirety.

[00214] Test compounds are employed as 50 millimolar (mM) solutions in dimethyl sulphoxide (DMSO). Ganciclovir®, Foscamet® and Cidofovir® are used as reference compounds. After addition of in each case 2 μΐ of the 50, 5, 0.5 and 0.05 mM DMSO stock solutions to 98 μΐ portions of cell culture medium in row 2 A-H for duplicate determinations, 1 :2 dilutions are carried out with 50 μΐ portions of medium up to row 11 of the 96-well plate. The wells in rows 1 and 12 each contain 50 μΐ of medium. In 150 μΐ of a suspension of l x lO⁴ cells (human prepuce fibroblasts [NHDF]) are pipetted into each of the wells (row l=cell control) and, in rows 2-12, a mixture of HCMV-infected and uninfected NHDF cells (M.O.I.=0.001-0.002), i.e. 1-2 infected cells per 1000 uninfected cells. Row 12 (without substance) serves as virus control. The final test concentrations are 250-0.0005 μΜ. The plates are incubated at 37° C./5% C02for 6 days, i.e. until all the cells are infected in the virus controls (100% cytopathogenic effect [CPE]). The wells are then fixed and stained by adding a mixture of formalin and Giemsa's dye (30 minutes), washed with double-distilled water and dried in a drying oven at 50° C. The plates are then assessed visually using an overhead microscope (Plaque Multiplier from Technomara).

[00215] The following data can be acquired from the test plates:

• CC5o(NHDF)=substance concentration in μΜ at which no visible cytostatic effects on the cells are evident by comparison with the untreated cell control; • EC5o(HCMV)=substance concentration in μΜ which inhibits the CPE (cytopathic effect) by 50% compared with the untreated virus control; and

• SI (selectivity index)=CC₅o (NHDF)/EC₅o (HCMV).

[00216] Compounds disclosed herein are expected to demonstrate activity in this assay.

Human Cytomegalovirus Xenograft Gelfoam® Model

[00217] The procedure is carried out as described in US 7,196,086, which is hereby incorporated by reference in its entirety.

[00218] Animals: 3-4-week old female immunodeficient mice (16-18 g), Fox Chase SCID or Fox Chase SCID-NOD or SCID beige, are purchased from commercial breeders (Taconic M+B, Jackson USA). The animals are housed under sterile conditions (including bedding and feed) in isolators.

[00219] Virus growing: Human cytomegalovirus (HCMV), Davis or AD 169 strain, is grown in vitro on human embryonic prepuce fibroblasts (NHDF cells). After the NHDF cells have been infected with a multiplicity of infection (M.O.I.) of 0.01-0.03, the virus-infected cells are harvested 5-10 days later and stored in the presence of minimal essential medium (MEM), 10% fetal calf serum (FCS) with 10% DMSO at 40° C. After serial ten-fold dilutions of the virus-infected cells, the titer is determined on 24-well plates of confluent NHDF cells after vital staining with Neutral Red.

[00220] Preparation of the Sponges, Transplantation, Treatment and Evaluation: Collagen sponges l x l x l cm in size (Gelfoam®; from Peasel & Lorey, order No. 407534; K. T. Chong et al, Abstracts of 39^th Inters cience Conference on Antimicrobial Agents and Chemotherapy, 1999, p. 439) are initially wetted with phosphate-buffered saline (PBS), the trapped air bubbles are removed by degassing, and then stored in MEM+10% FCS. l x l O⁶ virus-infected NHDF cells (infection with HCMV Davis or HCMV AD169 M.O.I. =0.01) are detached 3 hours after infection and added in a drop of 20 μΐ of MEM, 10% of FCS, to a moist sponge. About 16 hours later, the infected sponges are incubated with 25 μΐ of PBS/0.1% BSA/1 mM DTT with 5 ng/μΐ basic fibroblast growth factor (bFGF). For the transplantation, the immunodeficient mice are anaesthetized with Avertin or a ketamine/xylazine/azepromazine mixture, the fur on the back is removed using a shaver, the epidermis is opened 1-2 cm, unstressed and the moist sponges are transplanted under the dorsal skin. The surgical wound is closed with tissue glue. 6 hours after the transplantation, the mice can be treated for the first time (on the day of the operation, there is one treatment). The next days, over a period of 8 days, the mice are treated with substance orally three times a day (7.00 h and 14.00 h and 19.00 h), two times a day (8.00 h and 18.00 h) or once a day (14.00 h). The daily dose is, for example 3 or 10 or 30 or 60 or 100 mg/kg of body weight, the volume administered is 10 ml/kg of body weight. The substances are formulated in the form of a 0.5% strength Tylose suspension with 2% DMSO or a 0.5% strength Tylose suspension. 9 days after

transplantation and 16 hours after the last administration of substance, the animals are painlessly sacrificed and the sponge is removed. The virus -infected cells are released from the sponge by collagenase digestion (330 U/1.5 ml) and stored in the presence of MEM, 10% fetal calf serum, 10% DMSO at -140° C. Evaluation takes place after serial ten-fold dilutions of the virus -infected cells by determining the titer on 24-well plates of confluent NHDF cells after vital staining with Neutral Red. The number of infected cells or infectious virus particles (infectious centre assay) after the substance treatment compared with the placebo-treated control is determined. Compounds disclosed herein are expected to demonstrate activity in this assay.

Human Cytomegalovirus Cvtopathic Effect Reduction Assay

[00221] The procedure is carried out as described in Lischka et al, Antimicrob. Agents Chemother., 2010, 54(3), 1290-1297, and Reefschlaeger J. et al, 2001, J. Antimicrob.

Chemother. 48:757-767, which are hereby incorporated by reference in their entireties. In brief, the addition of 2 μΐ test compound of 50, 5, 0.5, 0.05, 0.005, and 0.0005 mM DMSO stock solutions to 100 μΐ cell culture medium in duplicates is followed by serial 2-fold dilutions in 96-well microtiter plates. Each well is supplemented with 150 μΐ of either a suspension of 1 ^χ 10⁴ NHDF cells mixed with cell-free HCMV (multiplicity of infection [MOI], 0.03) or a suspension of 1 ^χ 10⁴ to 3 ^χ 10⁴ HCMV-infected and uninfected NHDF cells (MOI, 0.001 to 0.002). Noninfected and nontreated cells serve as controls on each plate. Final compound concentrations range between 250 and 0.00005 μΜ. Plates are incubated for 6 to 7 days at 37°C or until the virus control reaches 100% CPE. A mixture containing 20% Giemsa stain (Merck) and 5% formalin solution (Merck) is added to the wells for fixation and staining. After extensive washing, plates are dried at 56°C followed by visual evaluation using an overhead microscope (plaque magnifier; Tecnorama Ziirich). Each assay is performed at least in triplicate, and standard deviations calculated. The assay plate data are used to calculate the EC50 (CPE-RA), i.e., the concentration of drug that inhibits the CPE by 50% compared with an untreated virus-infected control. Compounds disclosed herein are expected to demonstrate activity (e.g., by reducing cytopathic effects) in this assay. Human Cytomegalovirus Plaque Assay

[00222] The procedure is carried out as described in Lischka et al, Antimicrob. Agents Chemother., 2010, 54(3), 1290-1297, which is hereby incorporated by reference in its entirety.

[00223] NHDF cells (lxlO⁵ to 2xl0⁵) seeded in 24-well tissue culture plates are infected by inoculating 0.1 ml of serial log dilutions of a suspension of infected and uninfected cells. After a 16-h adsorption period, the cell culture supernatant is replaced by 1 ml of a methylcellulose (MC) overlay medium (0.5% MC-Dulbecco modified Eagle medium

[DMEM]-10% fetal calf serum). Cultures are incubated for 7 to 14 days. A mixture containing 20% Giemsa stain (Merck) and 5% formalin solution (Merck) is added to the wells for fixation and staining. Subsequently, plates are visually evaluated by counting plaques. Results are expressed as PFU per ml titrated cell suspension. Compounds disclosed herein are expected to demonstrate activity (e.g., by reducing plaques) in this assay.

Human Cytomegalovirus Mouse Xenograft Model

[00224] The procedure is carried out as described in Lischka et al, Antimicrob. Agents Chemother., 2010, 54(3), 1290-1297, which is hereby incorporated by reference in its entirety.

[00225] The in vivo antiviral activity of AIC246 may be assessed using a HCMV xenograft mouse model as described by Chong et al. (1999). Briefly, Gelfoam hemostyptic gelatin devices (Upjohn) are cut aseptically into 1-cm² pieces. These implants are soaked in NHDF cell culture growth medium (GM), and sponges are brought to 37°C in a CO2 incubator. NHDF cells are infected with cell-free HCMV strain Davis at an MOI of 0.03. After 4 h, cells are collected by trypsinization followed by centrifugation at room temperature for 10 min at 800 x g. Cells are resuspended in GM and counted using a hemocytometer. Each Gelfoam implant is seeded with a suspension of 1 x 106 infected cells by pipetting the cells onto the sponges. Human cells are allowed to adhere to the collagen sponges for at least 3 to 4 h at 37°C. To enhance vascularization of the implant, 250 ng recombinant human basic fibroblast growth factor (Calbiochem) is pipetted onto each implant 1 h prior to

transplantation. Mice (18 to 25 g body weight) are anesthetized, and the Gelfoam sponges are implanted subcutaneously in the dorsoscapular area. After transplantation, mice are randomized and grouped in -10 animals per treatment group. Starting 4 h after

transplantation, mice are treated once daily with the indicated compounds for nine consecutive days. Drugs are applied per os by oral gavage. Total administration volume is 10 ml/kg. Mice are sacrificed after 9 days of treatment, and the Gelfoam implants are removed and digested with collagenases (Calbiochem) at 37°C. After 2 to 3 h, human cells are recovered by centrifugation and resuspended in GM. Subsequently, the isolated cell suspensions are serially diluted and mixed with uninfected NHDF indicator cells and PFU are determined by plaque assays as described above. Virus titers determined from isolated cells are given as PFU/ml. Compounds disclosed herein are expected to demonstrate activity in this assay.

Clinical Study for the Treatment of Human Cytomegalovirus Infection in Kidney Transplant Patients

[00226] The procedure is carried out as described in Stoelben et al, Transplant

International, 2014, 27, 77-86, which is hereby incorporated by reference in its entirety. Compounds disclosed herein are expected to demonstrate efficacy in treatment of cytomegalovirus infection.

Clinical Study for the Prophylaxis of Human Cytomegalovirus Infection in Hematopoietic- Cell Transplantation Patients

[00227] The procedure is carried out as described in Chemaly et al, New Eng. J. Med., 2014, 370(90), 1781-1789, which is hereby incorporated by reference in its entirety.

Compounds disclosed herein are expected to demonstrate efficacy in preventing, at least to some degree compared to control, cytomegalovirus infection.

[00228] From the foregoing description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.

Claims

CLAIMS What is claimed is:

1. A compound of structural Formula la

(la)

or a salt thereof, wherein:

at least one of R1-R25 is deuterium or contains deuterium.

2. The compound, or a salt thereof, as recited in Claim 1 wherein R9 is hydrogen.

3. The compound, or a salt thereof, as recited in Claim 2 wherein R3-R5 and R22-R25 are hydrogen.

4. The compound, or a salt thereof, as recited in Claim 3 wherein R7 and Rs are hydrogen.

5. The compound, or a salt thereof, as recited in Claim 3 wherein four of R14-R21 are

hydrogen.

6. The compound, or a salt thereof, as recited in Claim 3 wherein six of R14-R21 are

hydrogen.

7. The compound, or a salt thereof, as recited in Claim 3 wherein R14-R21 are hydrogen.

8. The compound, or a salt thereof, as recited in Claim 7 wherein R7, Rs, and R14-R21 are hydrogen.

9. The compound, or a salt thereof, as recited in Claim 1 wherein compounds have structural Formula II:

(II)

or a salt thereof, wherein:

10. The compound, or a salt thereof, as recited in Claim 9 wherein R7 and Rs are hydrogen.

11. The compound, or a salt thereof, as recited in Claim 9 wherein R14-R21 are hydrogen.

12. The compound, or a salt thereof, as recited in Claim 9 wherein R7, Rs, and R14-R21 are hydrogen.

13. The compound, or a salt thereof, as recited in Claim 12 wherein Ri is -CD3.

14. The compound, or a salt thereof, as recited in Claim 12 wherein R2 is -CD3.

15. The compound, or a salt thereof, as recited in Claim 12 wherein Ri and R2 are -CD3.

16. The compound, or a salt thereof, as recited in Claim 12 wherein R6 is hydrogen.

17. The compound, or a salt thereof, as recited in Claim 9 wherein compounds have structural Formula III:

or a salt thereof, wherein:

R6 and R14-R21 are independently selected from the group consisting of hydrogen and deuterium; and

at least one of Ri, R2, R6, and R14-R21 is deuterium or contains deuterium.

18. The compound, or a salt thereof, as recited in Claim 17 wherein four of R14-R21 are hydrogen.

19. The compound, or a salt thereof, as recited in Claim 17 wherein six of R14-R21 are hydrogen.

20. The compound, or a salt thereof, as recited in Claim 17 wherein R14-R21 are hydrogen.

21. The compound, or a salt thereof, as recited in Claim 17 wherein R14-R21 are deuterium.

22. The compound, or a salt thereof, as recited in Claim 17 wherein R6 is hydrogen.

23. The compound, or a salt thereof, as recited in Claim 17 wherein R6 is deuterium.

24. The compound, or a salt thereof, as recited in Claim 22 wherein Ri is -CD3.

25. The compound, or a salt thereof, as recited in any of Claim 22 wherein R2 is -CD3.

26. The compound, or a salt thereof, as recited in any of Claim 22 wherein Ri and R2 are - CDs.

27. The compound, or a salt thereof, as recited in any of Claims 1-23 wherein Ri is -CD3.

28. The compound, or a salt thereof, as recited in any of Claims 1-23 wherein R2 is -CD3.

29. The compound, or a salt thereof, as recited in any of Claims 1-23 wherein Ri and R2 are - CD₃.

30. The compound, or a salt thereof, as recited in any of Claims 1-23 wherein at least one of R1-R25 independently has deuterium enrichment of no less than about 10%.

31. The compound, or a salt thereof, as recited in any of Claims 1 -23 wherein at least one of Ri-R.25 independently has deuterium enrichment of no less than about 50%.

32. The compound, or a salt thereof, as recited in any of Claims 1 -23 wherein at least one of Ri-R.25 independently has deuterium enrichment of no less than about 90%.

33. The compound, or a salt thereof, as recited in any of Claims 1 -23 wherein at least one of Ri-R.25 independently has deuterium enrichment of no less than about 98%.

34. The compound as recited in Claim 1 wherein the compound has a structural formula

selected from the group consisting of

78

79

81

82

83

84

91

92

94

35. The compound as recited in Claim 1 wherein the compound has a structural formula selected from the group consisting of

36. The compound as recited in Claim 1 wherein the compound has the structural formula:

or a salt thereof.

37. The compound as recited in Claim 1 , chosen from

or a salt thereof.

38. The compound as recited in Claim 1, wherein the compound is or a salt thereof.

39. The compound as recited in Cl is

or a salt thereof.

40. The compound as recited in Cl is

or a salt thereof.

41. The compound as recited in any one of Claims 34-40 wherein each position represented as D has deuterium enrichment of no less than about 10%.

42. The compound as recited in Claim 41 wherein each position represented as D has deuterium enrichment of no less than about 50%.

43. The compound as recited in Claim 41 wherein each position represented as D has deuterium enrichment of no less than about 90%.

44. The compound as recited in Claim 41 wherein each position represented as D has deuterium enrichment of no less than about 98%.

45. A pharmaceutical composition comprising a compound as recited in any one of claims 1- 44 together with a pharmaceutically acceptable carrier.

46. A method of treatment of a viral infection-mediated disorder comprising the

administration of a therapeutically effective amount of a compound as recited in any one of claims 1 -44 to a patient in need thereof.

47. The method as recited in Claim 46 wherein the disorder is selected from the group consisting of human cytomegalovirus infection, human cytomegalovirus infection in stem cell recipients, human cytomegalovirus infection in bone marrow transplant patients, human cytomegalovirus infection in kidney transplant patients, viral infection, herpes viridae infection, cytomegalovirus infection, human cytomegalovirus infection in AIDS patients, human cytomegalovirus infection in organ transplant patients, human cytomegalovirus pneumonitis, human cytomegalovirus encephalitis, gastrointestinal human cytomegalovirus infection, systemic human cytomegalovirus infection, human cytomegalovirus infection in neonates, human cytomegalovirus infection in infants, human cytomegalovirus infection in pregnant women, human cytomegalovirus infection in immune-suppressed patients, human cytomegalovirus infection in cancer, and human cytomegalovirus infection-mediated tumors.

48. The method as recited in Claim 47 further comprising the administration of an additional therapeutic agent.

49. The method as recited in Claim 48 wherein the additional therapeutic agent is an antiviral selected from the group consisting of ganciclovir, valganciclovir, foscarnet, cidofovir, and valaciclovir.

50. The method as recited in Claim 47, further resulting in at least one effect selected from the group consisting of:

a. decreased inter-individual variation in plasma levels of the compound or a metabolite thereof as compared to the non-isotopically enriched compound; b. increased average plasma levels of the compound per dosage unit thereof as compared to the non-isotopically enriched compound;

c. decreased average plasma levels of at least one metabolite of the compound per dosage unit thereof as compared to the non-isotopically enriched compound;

d. increased average plasma levels of at least one metabolite of the compound per dosage unit thereof as compared to the non-isotopically enriched compound; and

e. an improved clinical effect during the treatment in the subject per dosage unit thereof as compared to the non-isotopically enriched compound.

51. The method as recited in Claim 47, further resulting in at least two effects selected from the group consisting of: a. decreased inter-individual variation in plasma levels of the compound or a metabolite thereof as compared to the non-isotopically enriched compound; b. increased average plasma levels of the compound per dosage unit thereof as compared to the non-isotopically enriched compound;

52. The method as recited in Claim 47, wherein the method effects a decreased metabolism of the compound per dosage unit thereof by at least one polymorphically-expressed cytochrome P450 isoform in the subject, as compared to the corresponding non- isotopically enriched compound.

53. The method as recited in Claim 52, wherein the cytochrome P450 isoform is selected from the group consisting of CYP2C8, CYP2C9, CYP2C19, and CYP2D6.

54. The method as recited in Claim 47, wherein the compound is characterized by decreased inhibition of at least one cytochrome P450 or monoamine oxidase isoform in the subject per dosage unit thereof as compared to the non-isotopically enriched compound.

55. The method as recited in Claim 54, wherein the cytochrome P450 or monoamine oxidase isoform is selected from the group consisting of CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2, CYP2R1, CYP2S1, CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11, CYP4B1, CYP4F2, CYP4F3, CYP4F8, CYP4F11, CYP4F12, CYP4X1, CYP4Z1, CYP5A1, CYP7A1, CYP7B1, CYP8A1, CYP8B1, CYP11A1, CYP11B1, CYP11B2, CYP17, CYP19, CYP21, CYP24, CYP26A1, CYP26B1, CYP27A1, CYP27B1, CYP39, CYP46, CYP51, MAOA, and MAOB.

56. The method as recited in Claim 47, wherein the method reduces a deleterious change in a diagnostic hepatobiliary function endpoint, as compared to the corresponding non- isotopically enriched compound.

57. The method as recited in Claim 56, wherein the diagnostic hepatobiliary function

endpoint is selected from the group consisting of alanine aminotransferase ("ALT"), serum glutamic-pyruvic transaminase ("SGPT"), aspartate aminotransferase ("AST," "SGOT"), ALT/AST ratios, serum aldolase, alkaline phosphatase ("ALP"), ammonia levels, bilirubin, gamma-glutamyl transpeptidase ("GGTP," "γ-GTP," "GGT"), leucine aminopeptidase ("LAP"), liver biopsy, liver ultrasonography, liver nuclear scan, 5'- nucleotidase, and blood protein.

58. A compound, or a salt thereof, as recited in any one of claims 1-44 for use as a

medicament.

59. A compound, or a salt thereof, as recited in any one of claims 1-44 for use in the

manufacture of a medicament for the prevention or treatment of a viral infection-mediated disorder.