CA2372731A1 - Oligo-amine/oligo-carboxy sulfonamides - Google Patents

Abstract

The invention relates to new sulfonamide derivatives of aryl or heterocyclic compounds and to their pharmaceutically acceptable salts. The new derivatives of the invention can be obtained with chemical synthetic processes known in the art.
For example, they can be prepared by mono- or poly-condensation of a suitably substituted sulfonamide compound. The sulfonamide compounds of the invention are active, in particular, as inhibitors of carbonic anhydrase isoenzymes and therefore they can be useful in therapy, in particular in the ophthalmic field. In fact these compounds are able to decrease the intraocular pressure in glaucoma and to improve the protection of the retina. The new sulfonamide derivatives can be administered to humans in the form of different pharmaceutical preparations, in particular eye drops.

Description

OLIGO-AMINE/OLIGO-CARBOXY SULFONAMIDES
BACKGROUND OF THE INVENTION
Field of the Invention [0001) This invention is in the fields of chemistry and pharmacology, and relates to non-protein drug molecules that inhibit the enzyme carbonic anhydrase (CA).
More particularly, these water-soluble drugs can be used in eyedrops, to inhibit certain isozymes of the enzyme, in a manner that reduces pressure inside; the eye.
This is useful, in particular, for treating glaucoma patients.
Description of the Related Art [0002) Carbonic anhydrase enzymes are very widespread in nature, and are crucially important in allowing the body to handle carbon dioxide as a primary byproduct of metabolism and respiration. Within red blood cells, certain forms of the carbonic anhydrase enzyme rapidly cause a molecule of carbon dioxide to be combined with a molecule of water, to form carbonic acid (H2C03). This acid promptly dissociates and loses one of its hydrogen atoms, to form bicarbonate ions (HC03'). Since carbonic acid and bicarbonate are both highly soluble in water, this allows the blood to handle large quantities of carbon dioxide; in addition, the bicarbonate forms a useful buffering system, which can move interchangeably between carbonic acid, bicarbonate, and carbonate ions (C03"), depending on the acidity of the surrounding fluids; as such, it helps sustain a stable pH in the blood.

[0003) When bicarbonate ions in the blood reach the lungs, a different isozyme of carbonic anhydrase rapidly carries out the opposite reaction, and splits off molecules of carbon dioxide, which are then exhaled. This releases the water molecules as well; some of these water molecules are exhaled, and others are returned to the bloodstream for reuse.

[0004) Not all carbonic anhydrase enzymes work directly with bicarbonate ions;
instead, various other versions of carbonic anhydrase (different versions of an enzyme are often called "isozymes") carry out reactions in which a molecule of water is added to, or split apart from, various other types of molecules. Indeed, the main distinguishing feature of carbonic anhydrase enzymes is that they can carry out either or both of the following two classes of reactions: (1) they can attach a water molecule to one or more types of organic molecules (i.e., to molecules which contain at least one carbon atom);
and/or, (2) they can detach and remove a water molecule, from one or more types of organic molecules.

[0005] Since both of those two steps are essential to the metabolism of every type of microbe and organism, carbonic anhydrase enzymes play crucial roles in microbes and plants, as well as animals. Indeed, carbonic anhydrase enzymes are so important in nature that they evolved at least three independent and separate times, i:n "gene families" that are usually referred to as the alpha, beta, and gamma families. Vertebrates (including humans) have carbonic anhydrase genes that evolved within the alpha family.

[0006] Within vertebrates, at least 14 different "isozyme:>" of carbonic anhydrase have been identified, in various different tissues, and in various <iifferent locations inside cells. These different types are usually designated by Roman numerals; for example, isozymes CA-I, CA-II, CA-III, and CA-VII are found primarily :in soluble form, in the aqueous liquid within cells known as the cytosol; by contrast, C~~-IV, CA-IX, CA-XII, and CA-XIV tend to cling fairly tightly to membranes, and CA-V is a mitochondria) enzyme. Carbonic anhydrase from humans is often referred to by the "h" prefix, such as hCA.

[0007] Because of their importance in both nature and medicine, carbonic anhydrase enzymes, and drugs which can inhibit carbonic anhydrase activity, are discussed in nearly any textbook on biochemistry or physiology. They are also discussed in greater detail in various review articles, including Supuran & ~Scozzafava 2000, the contents of which are incorporated by reference, as though fully set forth herein.

[0008] Isozymes CA-II (two) and CA-IV (four) are important in this invention, since they are involved in the formation of the clear watery fluid that fills the eye. This fluid in the eye is often referred to, imprecisely, as either the "aqueous humor", or the "vitreous humor". More accurately, the "aqueous humor" is the fluid in the front part of the eye (i.e., in front of a focusing lens which is suspended near the center of the eye; this is the lens that becomes cloudy, in people suffering from cataracts), while the "vitreous humor" is the fluid in the rear (posterior) part of the eye, behind the focusing lens.

[0009] Aqueous and vitreous humors are formed inside the mammalian eye through a constant, slow, steady secretion of water, by certain cellular structures within the eye. These liquids slowly pass through the eyeball over a span of days or weeks, then they exit from the eyeball with the aid of several structures, including the "trabecular drainage meshwork", the "sinus venosus sclerae" (also called Schlemm's canal), and the uvea (also called the vascular tunic). With this slow yet constant inflow and outflow of water, the liquid inside the eyeball is slowly and gradually replenished.

[00010] Inside a healthy eye, the aqueous and vitreous humors work together to maintain a slight positive pressure. This fluid pressure is exerted against the membranes that surround and define the eyeball (the sclera and cornea), and it helps the eyeball sustain its generally spherical shape. That fluid pressure i.s usually referred to by the phrase "intra-ocular pressure", and by its acronym, IOP. It can be measured quickly and easily, by a machine which blows a slight puff of air against the exposed front surface of the eye and which measures the deflection of the eye in response to that puff of air.

[00011] In a healthy eye, the IOP is usually in a raaage of about 9 to about 15 mm Hg (i.e., millimeters of mercury; this is the standard way to express liquid pressures, and refers to the height of a column of mercury that would be supported inside a U-shaped tube, if the fluid pressure inside an eye pressed against the mercury in one tube, while the other tube was open to atmospheric pressure). Fo:r comparative purposes, people who suffer from glaucoma which is serious enough to drive them to a doctor because of deterioration in their vision often suffer from IOP levels in the range of 25 mm Hg. IOP levels of about 15 mm Hg can be regarded as maximum safe levels;
levels of about 12 or 13 can be regarded as better, safer, and preferred target pressure levels, for glaucoma treatments.

[00012] It has long been known that elevated fluid pressure inside the eye can gradually damage the nerve cells in the retina. This type of damage results from several interrelated factors, including the fact that elevated press~.wes in the watery liquid inside the eye will exert pressure on the walls of the capillaries that serve the nerve cells in the retina. Since capillary walls must be extremely thin, to allow adequate transfer of nutrients and waste products, they cannot resist elevated pressures. This potential problem is further aggravated by the fact that, because of evolutionary reasons, the capillaries that serve most of the retina (excluding a small region called the macula, in the center of the retina) are positioned on the front surface of the retina; in that location, those capillaries are directly exposed to the liquid pressure inside: the eyeball, and no protection for them is provided by the sclera or other structural layers.
Therefore, an increase in fluid pressure, inside the eye, will press against retinal capillary walls, in a manner which effectively squeezes them and reduces the amount. of blood that flows through them.

[00013] In the retina, any reduction in blood flow poses a very serious threat of permanent damage to neurons in the retina. Retinal neurons are extremely sensitive, and they must constantly generate vast numbers of rods and cones, which have short life-spans on the retinal surface and which play a crucial role in generating chemical changes in response to impinging light waves. Therefore, nerve cells in the retina will begin to die if blood flow through the retinal capillaries is hindered, even if only slightly, by elevated fluid pressures inside the eye.

[00014] A person suffers from a condition called "ocular hypertension", if his or her intraocular pressure (IOP) becomes elevated above normal levels. If not treated, ocular hypertension will lead directly to the disease called glaucoma, which is said to occur as soon as any damage or deterioration of the vision is noticed, or can be detected by diagnostic instruments. The terms "ocular hypertension" and "glaucoma" are used interchangeably by many people, including physicians, for l:wo reasons:
(r) the condition of ocular hypertension will lead directly to the disease of glaucoma; and, (ii) more patients will realize and grasp the seriousness of problem, and the need to work to control it and prevent its damage, if the more blunt and menacing; word "glaucoma" is used.

[00015] Glaucoma is a major cause of blindness, aand it poses very serious health, social, and economic problems. If elevated IOP is not treated fairly soon after it begins, a cascade of neurodegenerative events can begin to occur, which cannot be stopped, even with proper treatment, after it passes a certain stage.

[00016] Because of the fact that carbonic anhydrase (CA) isozymes play key roles in releasing water molecules from organic molecules, ir~ should not be surprising that carbonic anhydrase isozymes are involved in the secretion of aqueous and vitreous humor, the watery liquids that fill the eyeball. In particular, hCA isozymes II and IV
(which are located in cellular structures known as "ciliary processes") are believed to play important roles in those secretions inside the eye; however, other isozymes may also be involved, to a greater or lesser extent.

[00017] Accordingly, CA inhibitor drugs have been used for more than 40 years to treat glaucoma and ocular hypertension. This began in the early 1950's, when certain sulfonamide drugs, such as acetazolamide, were found to be potent and effective as CA inhibitors. However, when those drugs were taken orally, they caused unwanted side effects, arising from the fact that they non-selectively inhibited multiple CA
isozymes, including various CA isozymes that play important roles throughout the entire body.

[00018] In the early 1980's, a major breakthrough i.n the treatment of glaucoma was established, with the development of the first CA inhibitor drugs that could be applied directly to the eyes, in the form of eyedrops (reviewed in Maren 1995). For obvious reasons, this form of targeted and limited administration did not cause nearly as many side effects in other parts of the body. This breakthrough then led to the development of various improved sulfonamide drugs, including dorzolamide (Ponticello et al 1987} and brinzolamide (Silver et al, 2000a and 2000b), which are widely used today in clinical practice.

[00019] Dorzolamide, the older drug, is sold in a 2% w/w eyedrop formulation, under the brand name TRUSOPT~, by Merck & Company; it also is one of the ingredients in an eyedrop suspension called COSOPT~, which also contains timolol, a beta-adrenergic antagonist. Dorzolamide reportedly has some solubility in water;
however, since the salt is derived from a weak base and a strong acid, the resulting compound is relatively acidic. TRUSOPT~ eyedrops have a pH of about 5.6, and COSOPT~ eyedrops have a pH of about 5.65.

[00020] Brinzolamide is not soluble in water, so biinzolamide eyedrops must be formulated as a suspension of microfine particles. A 1 % w/v suspension, pH 7.5, is sold under the brand name AZOPT~, by Alcon Laboratories. 'The efficacy of dorzolamide and brinzolamide in lowering IOP are very similar, in the eyedrop formulations sold commercially (2% dorzolamide, and I % brinzolamide). Since more data are publicly available on dorzolamide, it is used and referred to herein as a "benchmark" drug; this means that the chemical and performance; traits of the new drugs disclosed herein were evaluated by comparing them to 2% w/v d~~rzolamide suspensions (several tests described below also included brinzolamide eyedrops, as an additional comparative control). Any references herein to comparisons against dorzolamide refer to comparisons against commercially available aqueous eyedrops containing 2% (by weight) dorzolamide.

[00021) Both dorzolamide and brinzolamide eyedrops suffer from serious drawbacks and limitations. One major problem centers around the fact that they are not sufficiently effective, in reducing IOP, to provide adequate protection against glaucoma.
Even when used exactly as prescribed, they typically cause a drop of only about 4 or 5 mm Hg in normal eyes, and about 6 to 7 mm Hg in eyes suffering; from glaucoma.
Therefore, in patients with IOP levels of about 20 mm Hg or higher, dorzolamide or brinzolamide typically cannot bring IOP levels down to fully safe; levels that do not pose a long-term risk of damage, and a second "adjunct" drug must usually be prescribed for such patients, for use along with dorzolamide or brinzolamide. Those adjunct drugs include timolol, a beta-adrenergic receptor blocker, sold by itself under the trademark TIMOPTIC~ (and also sold as a second ingredient in COSOPT~~) by Merck, and latanoprost, a prostaglandin analog, sold under the trademark XA,LATAN~ by Pharmacia-Upjohn. Timolol and latanoprost are both believed to act, at least in part, by increasing levels of liquid outflow from the eye; however, because of adverse side effects, they usually are prescribed only to patients who suffer from severe ocular hypertension, and/or patients who do not respond adequately to eyedrops containing dorzolamide or brinzolamide.

[00022) Besides the obvious additional cost of multi-drug regimens, many patients have a substantially greater tendency to fail to comply with prescribed therapy, if they must take several different drugs, each day, to control a single problem. This is due to several contributing factors, including (i) many patients simply cannot afford all the drugs; (ii) the need to take several drugs every day, for a single problem, is both physically and emotionally aggravating and irritating; and, (iii) local and systemic side effects tend to mount up even higher, when several drugs must be used to treat a single problem in a single part of the body, especially when the eyes are; involved.

[00023] The problem of adverse side effects caused by dorzolamide and brinzolamide is a major problem, because they both cause irntation, when applied directly to the eyes. Dorzolamide, the older of the two drugs, rep~~rtedly causes relatively high rates of unwanted side effects, at least some of which appear to be directly related to its acidity; these effects include stinging, burning, itching, and reddening of the eyes, blurred or cloudy vision, and bitter taste. The newer drug, brinzo:lamide (in 1 % eyedrops) reportedly caused less irritation than 2% dorzolamide eyedrops (Silver et al 2000a);
nevertheless, brinzolamide eyedrops still caused significant irritation and other adverse effects in large numbers of patients. Presumably, at least some of these adverse effects are caused or aggravated by the fact that brinzolamide is a suspension of fine particles, rather than a solution, and applying any form of particles directly to the front surface of the eye is likely to cause irritation and other adverse effects.

[00024] Any form of irritation is a very serious problem, when caused by eyedrops that must be applied directly to the eyes several times each day, to be effective.
Obviously, a patient will not receive the full benefit of a drug, if 'the patient does not use it as often as prescribed. Significant irntation to the eyes, if it occurs chronically, several times each and every day, will strongly discourage its use, especially if the adverse results of skipping dosages will not be manifested until years later. Among the millions of patients who must use these eye-irritating drugs, large numbers of patients simply will not use them, for the rest of their lives, with the frequency and consistency needed to stave off glaucoma.

[00025] A substantial part of the irritation caused by dorzolamide and brinzolamide arises from the fact that they have very low levels of solubility in water (as used herein, any references to "solubility" refer to solubility in water, or in other aqueous solutions such as buffered saline solutions). To increase their solubility (and/or their ability to stay suspended) in aqueous solutions, they typically are prepared as hydrochloride salts; however, that approach causes an increase in acidity, which aggravates the problems of stinging, burning, and reddening of tree eyes.
Accordingly, since the CA inhibitors disclosed herein have substantially higher levels of solubility in water, they can avoid the need for hydrochloride additives or other solubilizing or suspending agents.

[00026] To try to overcome the problems that limit: previous CA inhibitors, the Inventors herein commenced, some years ago, a program of attempting to synthesize other types of sulfonamide drugs that would be more soluble in water, without losing their efficacy as topical CA inhibitors that could be used, in eyedrops, by glaucoma patients. A number of novel drug classes which resulted from those efforts have been described in various published articles, including Scozzafava et ail 1999a and 1999b, Supuran et al 2000, and Ilies et al 2000. However, none of those drugs has been commercialized, and the need remains for improved candidate drugs which can perform well in human clinical trials, and which have various other desiravble traits.

[00027] In particular, a serious and pressing need exists for improved CAI
drugs which can be applied as eyedrops, and which will achieve any or all of three highly useful and desirable traits. The first desired trait is this: an improved topical CAI should be able to reduce IOP by greater levels than can be achieved by 2 %
dorzolamide or 1 brinzolamide eyedrops.

[00028] The second highly desirable trait for an improved topical CAI is this: it should have a longer duration of action than dorzolamide or brinzolamide. Those two drugs tend to have a relatively short duration of action, measured in only a few hours.
For example, in a standard animal test using albino rabbits with normal eye pressures, the IOP reduction provided by dorzolamide peaks at the first hour after administration, at about 4 mm Hg. By the time another hour has passed, IOP reduction has dwindled to only about 2 mm Hg, and after one more hour, only about 0.5 mm Hg reduction in IOP
remains.

[00029] It should be noted that dorzolamide and brinzolamide typically are prescribed for use three times per day. That frequency level is a compromise, in an effort to keep irntation levels tolerable. Since IOP returns to previous levels after only 2 or 3 hours, better and more consistent protection could be achieved if patients used those drugs 6 or 8 times each day.

[00030] It should be recognized that the damage inflicted by abnormally high IOP levels are a function of both pressure, and time. For ob~~ious reasons, the more _g_ time a neuron must spend under conditions of semi-starvation, where its needs are being only partially met, the more likely it is to suffer lasting damage, ~~r to die. Therefore, if an improved CA inhibitor drug in eyedrops can provide a longer and better duration of action, the amount of nutrition and sustenance that will be provided to retinal neurons, by improved capillary blood flow that results from lowered IOP levels, will be directly increased.

[00031] The third desired trait is this: an improved CA inhibitor drug should have a desirable and useful balance between water-solubility, and lipophilicity.
On one hand, this means that a desired candidate CA inhibitor compound should have sufficient water solubility to allow it to be dissolved in neutral or nearly neutral aqueous solutions, at desired therapeutic concentrations. Nevertheless, it also should be recognized that extremely high levels of water solubility are also undesirable, and some degree of lipophilicity is preferred, to maximize the desired IOP lowering effects of the drug in vivo. If a drug compound administered by eyedrops is totally soluble in water, it will tend to stay entirely dissolved in the aqueous and vitreous humors inside the eye, and will not contact and react with CA enzyme molecules associated with cells and tissues, as readily or effectively as a drug having sufficient lipophilicity to <;ause it to adhere to, and react with, biological molecules in cells and tissues that the drug contacts.

[00032] Accordingly, any references herein to terms such as water soluble, solubility, hydrophilic, etc., should be considered with regard to the fact that effective CA
inhibitor drugs should have a level of water solubility which achiieves a proper and useful balance as follows: (i) they can be dissolved at desired concentrations in aqueous eyedrops that are not acidic; and, (ii) after they enter the eye, they have sufficient lipophilicity to cause them to contact and react with carbonic anlrydrase enzymes in tissue structures.

[00033] Accordingly, one object of this invention is to provide an improved class of carbonic anhydrase inhibitor drugs, which have at least one, and preferably two, and ideally all three of the following traits, when compared to dorzolamide or brinzolamide: (i) sufficient solubility in water, so that they can b~~ fully dissolved in non-acidic aqueous eyedrops at a highly effective concentration; (ii) an ability to cause greater reductions in IOP than dorzolamide or brinzolamide; and, (iii) an ability to reduce IOP
_g_ for longer spans of time than dorzolamide or brinzolamide.

[00034] Another object of this invention is to proviide an improved class of eyedrops, which contain water-soluble CA inhibitor drugs in a relatively neutral and non-acidic formulation (with a pH within a range of about 6.8 to about 7.5), which will cause less eye irritation than acidic suspensions of insoluble CA inhibil:ors.

[00035] Another object of this invention is to disclose and provide improved water-soluble CA inhibitor drugs, which can reduce IOP in an effective manner.

[00036] Another object of this invention is to disclose and provide improved water-soluble CA inhibitor drugs, for use in eyedrops, which contain alkylamine-carboxy constituents.

[00037] These and other objects of the invention will become more apparent through the following summary, drawings, and description of the preferred embodiments.
BRIEF SUMMARY OF THE INVENTION

[00038] Improved drugs are disclosed which can potently inhibit two specific isozymes of carbonic anhydrase (CA), designated as CA-II and CA-IV.
In healthy eyes, these enzymes help replenish and renew the clear watery liquid that fills the eyeball. However, in patients suffering from glaucoma or ocular hypertension, CA-II and CA-IV aggravate the problem of abnormally high infra-ocular pressure (IOP).
Therefore, drugs that can inhibit CA-II and CA-IV, formulated in eyedrops, are used to treat glaucoma.

[00039] The improved CA inhibitors disclosed herein comprise at least one sulfonamide portion and a solubilizing portion, the solubilizing portion providing at least one alkyl segment, at least one secondary or tertiary amine nitrogen atom, and, preferably, at least one carboxy group. The solubilizing portions are referred to herein as "alkylamine-carboxy" (AAC) portions. Because of molecular weight and mobility considerations, AAC portions preferably have about 6 or fewer amines, and about 10 or fewer carboxy groups.

[00040] When suitable AAC reagents are bonded to sulfonamide reagents having suitable reactive groups, the reaction will form compounds referred to herein as AAC-sulfonamide compounds. As shown by the results in Table l and elsewhere herein, many of these AAC-sulfonamide compounds of the present invention have traits which will allow them to perform very effectively as therapeutic drugs, which can be formulated in non-acidic eyedrops that can reduce and control intraocular pressure in mammals, including humans.

[00041] The activity of any particular compound in the AAC-sulfonamide class as defined herein can be evaluated for such use, using screening tests as disclosed herein, including enzyme inhibition and solubility tests which can be carned out in vitro or in vivo in animal tests or human clinical trials.

[00042] Those candidate AAC-sulfonamide compounds which do have the desired combination of traits can provide three major advantages, when compared to prior art drugs used in CA inhibitor eyedrops, such as dorzolamide and brinzolamide.

[00043] First, the new AAC-sulfonamide drugs di~,closed herein generally have a much greater solubility in water at neutral or near neutral pH, than dorzolamide or brinzolarnide, which require eyedrops that are acidic to keep those prior art drugs dissolved or suspended. The AAC-sulfonamide drugs disclosed herein do not require acidifying agents in the eyedrops, and can be prepared, for example, as 2%
solutions in eyedrops having, preferably, neutral or near-neutral pH.

[00044] Ocular administration of solutions at a pH which differs from the pH of tears results in ocular discomfort and stinging (Tang et al., Optom.
Vis. Sci. 73:746-749, 1996; Hams, et al, Optom. Vis. Sci. 67:84-88, 1990). The pH of tears in normal individuals has been reported to be in the range of about 7.0 to about 7.5 (Yamada et al, Curr. Eye. Res. 16:482-486, 1997; Mogens, Acta Ophthalmologica 66:485-489, 1988;
Carney et al., Arch Ophthalmol 94:821-824, 1976). The pH zone of ocular awareness which produces discomfort has been reported to be exhibited in solutions having a pH
below 6.6 or above 7.8 (Adler's Physiology of the Eye, 7th ed., Nloses, R.A., ed., The C.V. Mosby Company, St. Louis, MO, 1981, pp. 21-22). Thus, ~~ pH range of from about 6.6 to about 7.8 has been referred to as the ocular comfort range (Harris et.
al., supra, 1990). Thus, oculax solutions containing the compounds of the present invention are, preferably, at a pH of from about 6.6 to about 7.8 and, more preferably, from about 7.0 to about 7.5. Since CA inhibitor eyedrops are normally prescribed three times per day for glaucoma patients, irritation caused by acidic eyedrops discourages use of those eyedrops, and many patients end up skipping dosages, which undercuts the efficacy these drugs in staving off blindness over a span of years. Accordingly, water-soluble CA
inhibitors which can be dissolved in non-acidic, non-irritating eyedrops at a pH within the comfort range will be used more frequently, reliably, and effectively.

[00045] Second, preferred compounds of the new ~3AC-sulfonamide drugs are more effective than dorzolamide or brinzolamide, in reducing IOP levels, although compounds which are equally effective or less effective than dora.zolamide or brinzolamide are also included within the scope of the invention. Dorzolamide or brinzolamide are not especially effective in reducing IOP; as a result, many glaucoma patients end up having to take additional drugs, such as timolol and/or latanoprost, in conjunction with dorzolamide or brinzolamide, to control their ocular pressures. In standard animal models, improved AAC-sulfonamide drugs as disclosed herein can reduce IOP levels by amounts that are several times higher than the best results that can be achieved by dorzolamide or brinzolamide.

[00046] Third, preferred compounds of new AAC-sulfonamide drugs reduce IOP for substantially longer periods of time than dorzolarnide or brinzolamide, although compounds which are effective for equal periods or shorter periods than dorzolamide or brinzolamide are also included within the scope of the invention. As indicated in FIG. 13, the IOP reduction provided by dorzolamide, in a standard animal model, peaks at 60 minutes, and drops off fairly quickly after that peak is passed, to less than half its peak value at two hours after administration. By contrast, even after 4 hours, many of the AAC-sulfonamide drugs as disclosed herein provide better IOP
reductions than dorzolamide was able to provide at its peak. Because retinal and neuronal damage is a time-dependent process, the amount of effective eye protection provided by a drug that can reduce IOP will depend much more heavily on two-dimensional "area under the curve" amounts, than on brief peak-level reductions that drop off' and disappear quickly.
Therefore, the longer-lasting AAC-sulfonamide drugs disclosed herein provide much better overall retinal protection than previously available drugs such as dorzolamide or brinzolamide.

[00047] Any one of these three results, by itself, would represent a very important and useful accomplishment. In fact a number of the A~~C-sulfonamide compounds have achieved all three of these results, simultaneou:;ly. Thus, the new AAC-sulfonamide drugs offer major improvements over the prior art.
BRIEF DESCRIPTION OF THE DRAWINGS

[00048] FIGURE 1 shows the chemical structures of seven alkylamino-carboxy (AAC) reagents designated by Arabic numerals 1 throu~;h 7 and by acronyms for the common names.

(00049] FIGURE 2 (illustrating (A) N-containing reactive groups, and (B) hydroxyl reactive groups) shows the chemical structures of various sulfonamide compounds, designated as compounds A through T, which were used as reagents in creating the AAC-sulfonamide compounds disclosed herein.

[00050] FIGURE 3 shows the structures of several representative AAC-sulfonamide compounds.

[00051] FIGURE 4 shows a general scheme for thE; synthesis of AAC-mono-sulfonamide compounds.

[00052] FIGURE 5 shows the application of the general scheme for the synthesis of AAC mono-sulfonamide compounds from AAC reagent 5.

[00053] FIGURE 6 shows a scheme for the synthesis of single-isomer AAC
mono-sulfonamide compounds from AAC reagent 5 with the sulfonamide bound to an end carboxyl group.

[00054] FIGURE 7 shows a scheme for the synthe~~is of single-isomer AAC
mono-sulfonamide compounds from AAC reagent 5 with the sulfonamide bound to the central carboxyl group.

[00055] FIGURE 8 shows a general scheme for thc; synthesis of AAC-bis-sulfonamide compounds via the corresponding dianhydride of A.AC reagents 4-6.

[00056] FIGURE 9 shows a general scheme for thc; synthesis of AAC-bis-sulfonamide compounds via the corresponding AAC reagent.

[00057] FIGURE 10 shows a scheme for the synthesis of AAC-sulfonamide compound 4M.

[00058] FIGURE 11 shows a scheme for the synthesis of AAC-sulfonamide compound 4S.

[00059] FIGURE 12 a scheme for the synthesis of AAC-sulfonamide compound 1 C.

[00060] FIGURE 13 shows the effect of topically administered sulfonamide inhibitors (2% water solutions/suspensions)on the I(~P of normotensive albino rabbits wherein curve 1 is dorzolamide (hydrochloride salt, pH 5.5);
curve 2 is brinzolamide (suspension, hydrochloride salt, pH 5.5); curve 3 is compound SMz (trisodium salt, pH 7.0); curve 4: compound Zn-SMZ (suspension, pH 7.0); curve 5:
compound 4N (trisodium salt, pH 7.0); curve 6: compound Zn-4lV (suspension, pH
7.0).
DETAILED DESCRIPTION OF THE INVENTION

[00061] The novel AAC-sulfonamide compounds of the present invention have improved aqueous solubility at physiological pH. These At~.C-sulfonamide compounds are effective and long-lasting IOP-lowering agents when formulated into neutral or near neutral pH aqueous eyedrops. These eyedrops carp be used to treat glaucoma and/or ocular hypertension without the painful side effects associated with the administration of the currently available drugs dorzolamide and brinzolamide.

[00062] The novel AAC-sulfonamide compounds ~;an be formed, for example, by creating bonds (such as amide or ester bonds) between AAC reagents and aromatic or heterocyclic sulfonamide reagents, as described below. The aromatic or heterocyclic sulfonamide portion of an AAC-sulfonamide compound can be called the "head," and the alkylamine-carboxy portion can be called the "tail."

[00063] In Figure 1 and throughout this text, the seven AAC reagents that were tested and screened using the procedures disclosed herein are designated, for convenient reference, by the numbers 1 through 7. These seven compounds have the structures shown in FIG. 1. The acronyms for these compounds ~~re also indicated in FIG.
1, for convenience. The names of these compounds are: compound l, iminodiacetic acid (IDA); compound 2, nitrilotriacetic acid (NTA); compound 3, ethylenediamine diacetic acid (EDDA); compound 4, ethylenediamine tetraacetic acid (EDTA); compound 5, diethylenetriamine pentaacetic acid (DTPA); compound 6, ethylene-bis(oxyethylene-nitrilo)-tetraacetic acid (EGTA); and compound 7, ethylene-diarnine-dihydroxyphenyl diacetic acid (EDDHA).

[00064] The AAC reagents of the present invention each contain: ( 1 ) at least one secondary or tertiary amine; (2) at least two carboxyl groups; and (3) at least one alkyl segment, preferably C 1 _2 alkyl, between an amine nitro;;en and a carboxyl group. Preferably, the AAC reagents comprise from 1 to about 6 amine nitrogens and from about 2 to about 10 carboxyl groups.

[00065] It also will be recognized by skilled artisans that various other types of linkages can couple the AAC portion with the sulfonamiide portion. It will further be recognized that alternate methods, in addition to those disclosed herein, can be used to make the AAC-sulfonamide compounds of the present invention.

[00066] The twenty different sulfonamide reagents. that were tested and screened using the procedures disclosed herein are designated by the letters A
through T.
Their structures are shown in FIG. 2, which is divided into two parts. FIG. 2A
shows the structures of sulfonamide reagents which contained a nitrogenous reactive group(such as amino, imino, hydrozino, etc.) while FIG. 2B shows the structure, of sulfonamide reagents which used a hydroxy reactive group (-OH). The chemical names for these compounds, and the name of the company from which they were purchased or the method by which each compound was synthesized, are provided below in Example 1 and Table 5.

[00067] All of the sulfonamide reagents shown in 1FIGS. 2A and 2B contain a sulfonamide group (-SOZNH2) attached as a pendant moiety to an aromatic ring structure, or an unsaturated heterocyclic ring structure. For convf,nience, these reagents are referred to herein merely as sulfonamide reagents.

[00068] The candidate CA inhibitor drugs that were created by reacting an alkylamine-carboxy reagent with a sulfonamide reagent are referred to herein as AAC-sulfonamides. Typical reaction conditions are described in detail in Examples 2-S, and are illustrated in FIGS. 5-8. Briefly, these reactions involved dis;;olving the sulfonamide reagent in anhydrous acetone, adding the AAC reagent, and adding a carbodiimide reagent (with the general chemical structure of R 1-N=C=N-R2, where R 1 and R2 are relatively small alkyl groups; most of the carbodiimide reagents 'used contained either isopropyl groups at both ends, or an ethyl group at one end, and a dimethyl-aminopropyl group at the other end (this 1-ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide compound is abbreviated as EDAC). Under suitable conditions, these types ~~f carbodiimide reagents will cause a carboxylic acid group to react and condense with either an amine group or a hydroxy group, in a manner which effectively removes a water rr~olecule (i.e., a hydrogen proton is removed from the amine or hydroxy group, and a hydro~xy group is removed from the carboxylic acid group). This effectively creates an amide bond (if an amine reactive group was used, as shown in FIG. 5) or an ester bond (if a hydroxy reactive groups was involved, as shown in FIG. 6). This reaction also converts the central carbon atom in the carbodiimide structure into a carbonyl structure, with an oxygen atom coupled to it by a double bond, and with the flanking nitrogen atoms connected by single bonds, thereby creating a substituted carbonyl-diamide, also known as a substituted form of urea, with a formula of R1-NH-CO-NH-R2, where the R1 and R2 groups will normally be the same groups that had been present on the carbodiimide reagent.
This condensation reaction was monitored for completion using thin-l;~yer chromatography (TLC), and typically required 8 to 16 hours. After it was completed, the resulting AAC-sulfonamide compound was purified by solvent removal, washing, and drying. If necessary to remove impurities, the solid was dissolved in 10 to 2 0 mL of potassium phosphate buffer, and the solution was passed through a high-performance liquid chromatography (HPLC) column. Impurities normally eluted more rapidly;
retention times were typically in the 5 to 10 minute range for impurities, and 15 to 20 minutes for the desired product.

[00069] Several AAC reagents also required protecaive groups, to prevent secondary amine groups (in reagents 1 and 3) or hydroxy groups (in reagent 7) from being altered by the condensation reactions. In these cases, modified reagents containing protective groups called "80C" (an abbreviation for tert-butyl-o:~cy-carbonyl) were used.
After the condensation reaction involving carbodiimide was completed, the BOC
groups were removed by using trifluoroacetic acid (TFA) under cold conditions. This procedure is described in more detail in Example 4, and is illustrated in FIG'. 7. The use of BOC
groups to protect potentially reactive substituents during a synthesis procedure is well-known in the art; many reagents can be purchased which already contain BOC
groups at desired locations, and methods of adding BOC groups to vulneralble sites on a reagent (such as by treating a reagent with t-butyl-oxycarbonyl-azide) are: also known.

[00070] Since 7 AAC reagents were reacted with 20 sulfonamides, 140 "mono-sulfonamide" compounds (containing a single sulfonamide residue, bonded to an AAC residue) were created. Example structures for three of these; mono-sulfonamide compounds are shown in FIG. 3, as compounds 1A, 6M, and 4S. Since their chemical names are long and cumbersome, these compounds are referred to herein by simple number-letter combinations; as an illustration, compound 6M was created by bonding a single molecule of AAC reagent 6 to a single molecule of sulfonamide reagent M. The enzyme-inhibition data generated by a number of these compounds are provided in Table 1 A (for relatively potent compounds) and Table 1 B (for generalhr less potent compounds); these tables are discussed in more detail below. For comparative purposes, the numbers in parentheses in Tables 1 A and 1 B indicate the potency of the sulfonamide reagent by itself, with no AAC portion bonded to it, in inhibiting the same CA
isozyme.

[00071] In a number of cases, "bis-sulfonamide" compounds were also created, by reacting a double-molar ratio of a sulfonamide reager.~t with an AAC reagent.
This created molecules having an AAC residue in the center, with two sulfonamide structures bonded to its opposite ends. These "bis" compounds are referred to herein by subscript notations such as compounds 6M2 and 4Sz, as shown in. FIG. 3 and as listed in Tables 1A and 1B. These compounds also can be referred to as di-sulfonamides or bi-sulfonamides; however, it should be noted that the "bis-" prefix tends to imply that the two sulfonamide groups are bonded to opposite ends of the AAC reagent, rather than having both sulfonamide groups bonded to two adjacent carboxy groups at one end of the AAC molecule. The "bis" arrangement results from a method of using two dianhydride rings during synthesis, as discussed in more detail in Example S, and illustrated in FIG. 8.
This approach gives the resulting "bis" molecules a type of symmetry that is shown for compounds 6M2 and 452, in FIG. 3, and offers one method of preventing and avoiding the formation of isomeric mixtures, which are likely to be less desirable than isomerically pure forms, as discussed below. However, it should be recognized that various di-sulfonamide compounds which are not "bis" compounds as defined above can also be created (including compounds that are in isomerically pure form;. by various means, if desired, and are included within the claims.

[00072] Twenty different sulfonamide reagents were used to synthesize the novel AAC-sulfonamide compounds. Throughout this text they ~~re designated by the letters A through T, and their names are as follows: A, 2-amino-benzenesulfonamide; B, 3- amino-benzenesulfonamide; C, 4- amino-benzenesulfonamide; D, 4-hydrazino-benzenesulfonamide; E, 4-aminomethyl- benzenesulfonamide; F, 4-(2-aminoethyl)-benzenesulfonamide; G, 3-fluoro-4-amino-benzenesulfonamide; H, 3-chloro-4-amino-benzenesulfonamide; I, 3-bromo-4-amino-benzensulfonamide; J, 3-fluoro-4-amino-benzenesulfonamide; K, 4,5-dichloro-6-amino-benzene-1,3-disul.fonamide; L, 6-chloro-4-amino-benzene-1,3-disulfonamide; M, 5-amino-1,3,4-thiadiazol-2-sulfonamid; N, imino-4-methyl-2-sulfonamido-82-1,3,4-thiadiazoline; O, S-(2-arninoethylcarboxamido)-1,3,4-thiadiazol-2-sulfonamide; P, 6-amino-benzothiazol-2-sulfonamide; Q, 6-hydroxy-benzothiazol-2-sulfonamide; R, 6-(2-hydroxyethyloxy)-benzothiazol-2-sulfonamide; S, 4-hydroxymethyl-benzenesulfonamide; and T, 4-(2-hydroxyethyl)-benzenesulfonamide.
The structures of the twenty sulfonamide reagents are shown in I~ IG. 2A
(sulfonamide reagents A-P) and FIG. 2B (sulfonamide reagents Q-T). FIG. 2A shows the structures of sulfonamide reagents with a N-containing reactive group (amino.~imino/hydrazino) and FIG. 2B shows the structures of sulfonamide reagents which contain a hydroxyl reactive group.

[00073] Sulfonamide reagents A-T contain at least one sulfonamide group (-SOzNH2) attached to an aromatic or heterocyclic single-ring or double-ring structure.
Therefore, these reagents can be referred to as aromatic sulfonamides when the sulfonamide group is attached to an aromatic ring structure or as heterocyclic sulfonamides when the sulfonamide is attached to a heterocyclic ring structure.

[00074] As a large number of AAC-sulfonamide compounds are within the scope of the invention, we have abbreviated the names herein by using both a numeral designating the AAC reagent from which they were derived as well as a letter designating the sulfonamide at which the AAC moiety has been attached. Fo-.r example, 1 A
is the monoamide of iminodiacetic acid 1 with orthanilamide A; 6M is the monoamide of EGTA 6 with S-amino-1,3,4-thiadiazole-2-sulfonamide M; 6M2 is the bis-amide of EGTA 6 with the sulfonamide M; 4S is the monoester of EDTA 4 with 4-(2-hydroxymethyl)-henzenesulfonamide S. An abbreviation 1(A-T)includes the monoester/monoamide compounds created by reacting AAC reagent 1 with sulfonamide reagents A through T. An abbreviation (4,6)(A-T)2 includes the di-ester/di-amide compounds created by reacting AAC reagents 1 and 6 with sulfonamide reagents A
through T.
SYNTHESIS OF MONO-SULFONAMIDE A,AC-SULFONAMIDE COMPOUNDS

[00075] Synthesis of 140 "mono-sulfonamide" AAC-sulfonamide compounds was achieved by reacting one of the seven AAC reagents 1 to 7 with one of the 20 sulfonamides A to T. Example structures for three of thesc; mono-sulfonamide compounds are shown in FIG. 3, as compounds 1 A, 6M, and 4S.

[00076] A general scheme for the synthesis of mono-sulfonamide AAC-sulfonamide compounds is shown on FIG. 4. AAC reagents 2,4,:> and 6 were used in the synthesis reactions without prior modification, while reagents 1, 3 and 7 were modified by the addition of protective tert-butyl-oxycarbonyl (BOC) groups to the corresponding secondary amino and hydroxy groups present in these reagents. ~Che bulky BOC
groups displace the hydrogen in the corresponding amino (in AAC reagc;nts 1 and 3) or hydroxyl groups (in reagent 7) thus preventing sulfonamides A-T from reacting with the BOC-protected group. Thus, a single BOC-protective group was added to the amino group of reagent 1, two BOC-protective groups were added to the two amino groups of AAC
reagent 3, and two BOC-protective group were added to the two hydroxyl groups of AAC
reagent 7. Methods of adding BOC-protective groups are described in the literature and are well known to those skilled in the art. For example, one method of adding a BOC-protective group is by treating the corresponding AAC reagent with tert-butyl-oxycarbonyl-azide.

[00077] The general procedure for preparation of mono-sulfonamide AAC-sulfonamide compounds ( 1-7)(A-T) was as follows. 1 mMol of sulfonamide A-T
was dissolved/suspended in 25 ml of anhydrous acetone and then trez~ted with an equimolar amount (1 mMol) of AAC reagent 2,4,5 or 6, or BOC-protected .AAC reagents 1,3 or 7.
An amount of 190 mg (1 mMol) of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDCI~HCI)(or an equivalent amount of 1,3-di-isoproyl-carbodiimide) was then added and the reaction mixture was magnetically stirred at room temperature for 15 minutes.

[00078] When EDChHCI is added to the corresponding sulfonamide/AAC
reagent mixture, a hydroxyl group is removed from carboxylic acid, a hydrogen is removed from the reactive group of the sulfonamide A-T, and a covalent bond is formed between the AAC portion (1-7) and the sulfonamide portion (A-':('). Because sulfonamides A-T have different reactive groups (i.e. amino, imi:no, hydrazino or hydroxyl reactive group) which react with the AAC reagents 1-7., two different types of covalent bonds axe formed depending on the type of sulfonamide reactive group involved. Sulfonamides A-P (shown on FIG. 2A) have a N-containing reactive group (amino/imino/hydrazine) and bond to the AAC reagents 1-7 via an amide bond (as shown for AAC-sulfonamide compounds 1A a.nd 6M on FIG. 3), while sulfonamides Q-T
(shown on FIG. 2B) have a hydroxyl reactive group and bond to the AAC reagents via an ester bond (as shown for AAC-sulfonamide compound 4S on FIG. 3).

[00079] After the reaction mixture was stirred for 1. 5 minutes at room temperature, 30 wL (2mM) of triethylamine were added (TLC control) and stirnng was continued at 4°C until thin layer chromatography indicated that the desired reaction was complete; this usually required 8-16 hours. The solvent was evaporated in vacuo. The residue was taken up in 5 ml of ethyl acetate, poured into 5 mL of 5% solution of sodium bicarbonate, brought to pH 7 with 1 M HCI and extracted with ethyl acetate.
The combined organic layers were dried over sodium sulfate and filtered, and the solvent removed in vacuo.

[00080] In the cases when BOC-protected AAC reagents 1,3 or 7 were used, the obtained oils were either directly used in the de-protection step, or the intermediates were first purified by preparative HPLC as described below. The removal of the protecting groups has been performed as described in the literature.
The BOC-protected intermediate was dissolved in 20 mL of methylene chloride (CHZC12), treated with 4 mL of trifluoroacetic acid (TFA) and stirred for 10 min at 0°C.
The solvent was removed in vacuo and the residue concentrated from water twice to remove the excess TFA, giving thus the unprotected derivative as a colorless syrup.

[00081] AAC-mono-sulfonamide compounds were: separated from bis-sulfonamide compounds and other reaction by-products by preparative HPLC, using C 18 reversed-phase ~,-Bondpack or Dynamax-60A (25x250mm) columns, with elution buffer containing 90% acetonitrile, 8% methanol and 2% phosphate buffer, pH 7.4, at mL/min. Impurities (i.e. bis-sulfonamide compounds and reaction by-products) normally eluted within 5-10 min, whereas desired mono-sulfonamide compounds eluted after 15-20 min.

[00082] When sulfonamides A-T were reacted with AAC reagents 1-7 in ratio 1:1, as described in the preceding paragraphs, the predominant product was substituted with only one sulfonamide A-T moiety via an amide or ester bond, i.e. mono-sulfonamide compounds were the predominant reaction product. However, a small amount of the reaction product was substituted with two sulfonamide A-T
moieties, i.e.
some bis-sulfonamide compounds were formed in the reaction as. well. Due to the difference in molecular weight between mono-sulfonamide and bis-sulfonamide compounds, they were separated by preparative HPLC.

[00083] It should be recognized that AAC reagents. 1-4 and 6 and 7 (i.e. all AAC reagents other than 5) are entirely symmetric with respect to the placement of their carboxyl(-COOH) groups. Because these AAC reagents are synvnetric, all the carboxyl groups in each of these molecules are equivalent. Therefore, a single product was obtained with no differing isomers when these AAC reagents were reacted with sulfonamides A-T in ratio 1:1 under the conditions described above.

[00084] AAC reagent 5 (DTPA) differs from the other AAC reagents used in that it has an additional carboxyl group attached to the central nitrogen atom. (For convenience, this carboxyl group will be called "central" carbox:~ group, and the carboxyl groups attached to the outer nitrogens will be called "end" carbo:~cyl groups.) As a result, when AAC reagent S was reacted with sulfonamides A-T in ratio 1:1 under the conditions described in the preceding paragraphs, the reaction product(which is shown on FIGS) was a mixture of two distinct isomers in an estimated ratio of 4:1. Thus, an estimated 20% of the reaction product had the sulfonamide moiety bound t~~ the central carboxyl group, and an estimated 80% of the reaction product had the sulfbnamide moiety bound to an end carboxyl group. These two-isomer mixtures of mono-sulfonamide compounds formed from AAC reagent S were used in the in vitro enzyme inhibition tests and in the in vivo animal tests described below.

[00085] Isomeric mixtures of the two mono-sulfonamide compounds formed from AAC reagents having non-identical carboxy groups, as well as substantially isomerically pure preparations, are within the scope of the invention. If desired, such substantially pure single-isomer preparations of mono-sulfonamide compounds can be prepared as well. By substantially pure, it is meant that one isomer largely predominates in the preparation, and, preferably the preparation is substantially free of other isomeric forms of the compound. The pure isomers are, preferably, present in an amount greater than 80% (w/w) with any other isomers being present in an amount less than 20%. More preferably, the pure isomers constitute at least about 90% of the ~~omposition, still more preferably, at least about 95% and most preferably, at least about 98% up to about 100%.

[00086] Substantially pure isomers can be prepared by a number of methods. To illustrate two approaches that could be used to accomplish that type of goal, FIGS. 6 and 7 depict synthetic schemes that could be used, if desired, to create isomerically pure mono-sulfonamide compounds from DTPA. These synthetic schemes are described in more detail in Examples 12 and 13. Briefly, FIG. 6 shows a method of creating a mono-sulfonamide with a single sulfonamide-plus-ring; residue bonded to one of the end-connected carboxy groups of DTPA, by first placing a. methyl protective group on the center carboxy group of DTPA dianhydride. The dianhydiide compound is then hydrated, to generate four free carboxy groups at the ends of the methyl-ester intermediate. This methylated and hydrated intermediate is then reacted with a single molar equivalent of the sulfonamide reagent, in the presence of a carbodiimide reagent, causing the sulfonamide reagent to react with one of the four end-connected carboxy groups, all of which are structurally equivalent to each other. The; methyl protective group can then be removed from the center carboxy group, by using a "condensate polishing" resin in mild acid. Any undesired di-, tri-, or tetra-sulfonamides can be removed from the desired mono-sulfonamide product, by chromatography, electrophoresis, or any other suitable purification method, based on the large differences in their molecular weights.

[00087] FIG. 7 shows a method of creating a mono-sulfonamide with a single sulfonamide residue bonded to one of the "end-connected" carboxy groups, by reacting two molecules of a halogenated di(methylester)amine derivative with aminoacetate, thereby creating a DTPA structure with methyl protective groups on all four of the end carboxy groups. This reagent, having a single unprotected carboxy group in the center, is then reacted with a single molar equivalent of the sulfonamide reagent, to create a tetra-methyl-protected mono-sulfonamide intermediate. This intermediate is then passed through a suitable type of ion exchange resin (such as onf; of the "condensate polishing" DOWEX resins sold by Dow Chemical Company)) wader acidic conditions to remove the four methyl protective groups from the end-connected carboxy groups, leaving the final mono- sulfonamide product.
SYNTHESIS OF BIS-SULFONAMIDES

[00088] Synthesis of numerous A,AC bis-sulfonamide compounds using AAC reagents 4-6, was achieved utilizing the general scheme sh~~wn in FIG.B.
Bis-sulfonamide AAC-sulfonamide compounds were synthesized via the corresponding dianhydride of AAC reagents 4-6. An amount of 15 mMol of the corresponding dianhydride was added to a solution of 30 mMol of sulfonamide A-T dissolved in mL of anhydrous dimethylformamide (DMF). The mixture was magnetically stirred at room temperature for 4 hours, then the reaction mixture was poured in 300 mL
of methylene chloride (CHZC12) and the obtained solid was filtered and thoroughly washed with methylene chloride and then with acetone. HPLC was nece:~sary in some cases and was done by elution with potassium phosphate buffer-MeCN 2:1 (v/v) at 10 mL/min, on a reversed-phase C ~ 8 Bondapack column.

[00089] The dianhydride method of synthesis results in the formation of "bis"-sulfonamide AAC-sulfonamide compounds such as compounds 4S2 and 6M2 in FIG. 3. If desired, these compounds can be referred to as di-sulfonamides or bi-sulfonamides; however,the "bis-" prefix is preferred, since it tends to imply that the two sulfonamide groups are bonded to the opposite ends of the A.AC reagent rather than having both sulfonamide groups bonded to two adjacent carboxyl groups at one end of the AAC moiety.

[00090] The dianhydride method of synthesis results in the formation of isomerically single products, which are AAC bis-sulfonamide compounds with no isomeric forms. The two sulfonamide moieties are bound to two end carboxyl groups as shown on FIG. 3 and 8, and no sulfonamide moiety can bind to the central carboxyl group of a compound such as AAC reagent 5. Drawing a sulfonamide group at an "upper right" versus "lower right" (compare compounds 4S2 and 6M2 in FIG. 3) position makes no difference, since the AAC-sulfonamide compounds contain at least one single bond which allows free rotation of the parts of the molecule with respect to each other.
Therefore, compound 6M2 could be drawn with the right-end sulfonamide group shown in either the upper or lower right position, and either drawing would represent exactly the same compound.

[00091] Bis-sulfonamide compounds prepared from AAC reagents 1-3 and 7 can be created when the general scheme shown in FIG. 9 is utilized. This scheme is nearly identical with the general reaction scheme for the synthesis of mono-sulfonamide AAC-sulfonamide compounds shown in FIG. 4. The difference between the two schemes is that when bis-sulfonamide compounds in FIG. 9 are synthesized, the amount of the corresponding sulfonamide reagent added to the reaction mixturf; will be doubled compared to the amount that was added when the mono-sulfonamide sulfonamide compounds were synthesized. The bis-sulfonamide compounds will be the predominant product and can be separated from the reaction by-products such as mono-sulfonamide compounds by preparative HPLC due to the differences in molecular weight.

[00092] Substantially pure isomers can also be obtained by separation of individual isomers from mixtures, using chemical derivatization, chromatography and/or electrophoresis methods. T'he chromatography can comprise any suitable chromatography method, such as high pressure liquid chromatography (Dan, N., (Janesan, R., Flood, K.G., Tsai, D. and Reif, V.D. Determination of enantiomers in a synthetic argininal peptide using capillary zone electrophoresis and high-performance liquid chromatography. J. Chromatography A 891:115-127 (2000)), low pressure liquid chromatography (Sreenivas Rao, D., et al., LC separation of ortho and meta isomers of celecoxib in bulk and formulations using a chiral column. J Pharm. Biomed.
Anal. 25:
21-30 (2001) or thin layer chromatography (Thomas, M.H., Epstein, R.L., Ashworth, R.B. and Marks, H., Quantitative thin layer chromatographic multi-sulfonamide screening procedure: collaborative study. J. Assoc. Ofd'' Anal. Ch.em. 66: 884-892 (1983).

Electrophoresis can be gel electrophoresis or capillary electrophoresis (Driouich, R., Takayanagi, T., Oshima, M. and Motomizu, S., Separation and determination of n-alkylamines and histamine by capillary zone electrophoresis using salicylaldehyde-5-sulfonate as a derivatizing reagent. J. Chromatogr. A. 934: 95-103 (2001) [00093] It should be recognized that, except for D'CPA (reagent #5), all of the numbered AAC compounds shown in FIG. 1 are entirely symmetric, with respect to the placement of their carboxyl (-COOH) groups. The symmetric; AAC compounds (i.e., the AAC compounds other than DTPA) therefore form a distinct group. Because these compounds are symmetric, the carboxyl moieties are equivalent in each molecule.
Therefore, a single species results from mono-sulfonamide derivativation of one of these AAC compounds. A general scheme for synthesizing mono-sulfonamide compounds is shown in FIG. 5, and is discussed in more detail below.

[00094] Similarly, when bis-sulfonamide compounds such as 6M2 or 4S2 (also shown in FIG. 3) are created (as shown in FIG. 6, discussed in more detail below), the positioning of a sulfonamide group at an "upper right" versus "lower right" position makes no difference, since the chain which connects the right end of the molecule to the left end of the molecule contains one or more single bonds. Any single bond will allow the two ends of a molecule to rotate, with respect to each other, ~cround that single bond.
As can be seen in FIG. 3, compound 6M2 contains 9 single bonds between the two tertiary amines at the ends of the straight chain, and compound 4S2 contains 3 single bonds. Therefore, compound 6M2 could be drawn with the right-end sulfonamide group shown in either the upper or lower right position, and either drawing would represent exactly the same compound. Similarly, compound 4S2 could be drawn with either of the two sulfonamide groups in either the upper or lower position; there are four different ways to draw that compound, and all four drawings would show the same isomer of the same compound.

[00095] Isomer formation can be reduced substantiially by using a "di-anhydride" version of DTPA, which is also shown in FIG. 1 as an unnumbered compound, labelled "DTPA dianhydride". DTPA anhydride was tested as an AAC
reagent, as described in Scozzafava et al 2001, and the resulting .AAC-sulfonamide compound performed relatively well in animal tests. Accordinghr, DTPA
anhydride can be used as a specific reagent to avoid the production os isomeric mixtures.

[00096] Bis-sulfonamide compounds can be prepared without generating a mixture of isomers using the dianhydride and a double molar ratio of sulfonamide reagent to anhydride.

[00097] The AAC reagents shown as compounds 1l-7 each contain: (1) at least one secondary or tertiary amine nitrogen atom (i.e., a nitrogen atom which is positioned internally, and which is coupled to at least two carbon or other non-hydrogen atoms, as distinct from a "primary" amino group (-NHZ) which is at the end of a chain or group; and, (2) at least two carboxylic groups. Accordingly, these compounds can be regarded as "oligo-carboxy" reagents, and most of these reagent;> also qualify as oligo-amine reagents as well (although AAC reagents 1 and 2 contain .only I amine nitrogen).
As used herein, the prefix "oligo" indicates two or more. While there is no specific upper limit to the number of amine nitrogens or carboxy groups or residues which can be present in an AAC reagent, it is anticipated (because of molecul~~r weight and motility considerations, discussed in more detail below) that reagents having from 1 to about 6 amine nitrogens, and 2 to about 10 carboxy groups are preferred [00098) It should also be noted that some types of AC reagents contemplated for use as disclosed herein may contain only one carboxy group.
Instead of using a carboxy group to react with an amine or hydroxy group on the sulfonamide reagent (thereby forming an amide or ester bond, as shown in Fh3S. 3 and 5-7), the AC
reagent can contain the amine or hydroxy group, and the sulfonamide reagent can contain the carboxylic acid reactive group. This approach would enable the same type of condensation reaction, creating the same type of amide or ester bond, oriented in the opposite direction.

[00099] It also will be recognized by sl~cilled chemists that: (i) various other types of reactive groups could be used on either or both reagents., so long as the chosen combination of reactive groups will react with each other under suitable conditions to form a stable bond between the sulfonamide-and-ring component, and the amine-carboxy component; and, (ii) alternate pathways to creating such compounds can also be developed, such as by bonding a sulfonamide reagent that has not yet been bonded to a ring structure, to an AAC-plus-ring reagent that already contains a desired AAC "tail"

component bonded to an aromatic or unsaturated heterocyclic ring structure.

[000100] It is generally anticipated that AAC-sulfonamide compounds having relatively small molecular sizes, as indicated by molecul~~r weights generally in a range of less than about 1000 daltons, are likely to perform more; effectively than other potential candidate molecules having molecular weights substantially higher than about 1000 daltons, due to factors such as the reduced motility of larger and heavier molecules in aqueous solutions.

[000101] Preferrably, the AAC compounds of the pxesent invention include the following:
(i) from 2 to about 6 carbon atoms, between "end" amine nitrogen atoms in the chain that separates carboxy groups or anhydride rings from each other;
(ii) from 1 to about 3 oxygen atoms, in ether, ester, thioester, or amide linkages, or possibly in ketone or other carbonyl or imine linkages;
(iii) from 1 to about 5 nitrogen atoms, in secondary or tertiary amine structures.

[000102] It is further assumed that one or more oxygen atoms present in these compounds might be replaceable by sulfur atoms, secondaay amines, N-methyl groups, or similar groups, provided that the stability of the resulting AAC
reagent is not seriously reduced.

[000103] It is further assumed that, to avoid unwanted isomer mixtures, preferred AAC reagents generally should be symmetric, as occum with compounds 1, 3, 4, and 7 as shown in FIG. 1, or possibly in a radial manner around a central atom, such as the central nitrogen atom shown in compound 2.

[000104] The metal complexes of some of the above-mentioned sulfonamides were prepared as well. Zinc, copper and aluminum metal complexes were prepared with 4M2, SN2 and 6M2 as ligands. Zinc complexes were also prepared with 4C2, 4E2, 4F2, 4N2, SC2, SE2, SF2, SN2, 6C2, 6E2, 6F2 and 6N2 as ligands.

[000105] The various metal complexes were prepared as follows. 2 mMol of the corresponding bis-sulfonamide compound were treated with the stoichiometric amount of 1 M NaOH to obtain the disodium carboxylate salt. The obtained solution was treated with ImMol of ZnCl2 (to prepare Zn(II) complex)/ CuS04 (to prepare Cu(II) complex)/ Al2(S04)3 (to prepare Al(III) complex) in 5 mL of water, maintaining the pH
at 6.5. The reaction was monitored by HPLC, on a stationary phase of Lichrosphere 100 RP-18.5 um, with a 250x4 mm column packed by E. Merck, at CEO°C.
Isocratic elution with premixed mobile phase ( 1 g octylamine was added to 100 mL of acetonitrile mixed with 900 mL of water) was performed. The eluent was buffered 'with phosphoric acid, maintaining the pH at 6, the flow rate was of 1.5 mL/min. After 4 h the solution was loaded onto an Amberlite XAD 1600 resin column (250 mL) and eluted with MeCN/water ( 1:10, v/v). The fractions containing the complex vrere evaporated to give a white solid of the complex with an overall yield of 95 %.
CHEMICAL STRUCTURES

[000106] Preferred compounds of the present invention axe carbonic anhydrase inhibitory compounds or salts thereof. Each compound comprises at least one sulfonamide group attached to an organic ring, and a tail portion to embody the formula ZL", where n can be I or 2. Thus the compounds can be mono-substituted or bis-substituted.

[000107] In these compounds, Z is a tail portion, and L is a ring with at least one sulfonamide attached. Z is selected from the group consisting of:
O O

R, II R2 N R3 II

R

O O
R, ~) R2 R3_-II
\N R' X' Rs N\
R5 / Rs R~ ~ ~ R2 R3 Rs x2 Rio Xs R»
Rs ~Rs and R~ C R2 R3 C
R12-Xa R~3-Xs Rya Xs R~s R5 / Rs R' is hydroxy when n is 1;
R' is a bond when n is 2;
RZ and R3 are independently selected from the group consisting of a bond, -CHZ-, and -(CH2)2-;
RS and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C ~ _2-alkyl, wherein the C ~ _2-alkyl i~.s optionally substituted with carboxy;
X' is selected from the group consisting of a bond, -O-, -S-, and -N(RI6)-;
X~ is selected from the group consisting of-O-, -S-, and -N(R")-;
X3 is selected from the group consisting o~F-O-, -S-, and -N(R'x)-;
X4 is selected from the group consisting o~F-0-, -S-, and-N(R'9)-;
XS is selected from the group consisting o~F-O-, -S-, and N(RZ°)-;
X6 is selected from the group consisting oaf -0-, -S-, and N(RZ' )-;
R4, R'6, R", R'g, R'9, RZ° and RZ' are independently selected from the group consisting of hydrogen, carboxy, and CI_2-alkyl, wherein the C1_2-alkyl is optionally substituted with carboxy;
R' and Rg are independently selected from the group consisting of C~_5-alkyl, wherein the sum of the carbon atoms in R7 and Rg is from 2 to 6;
R9, Rl° and Rl l are independently selected from the group consisting of C1.~-alkyl, wherein the sum of the carbon atoms in R9, R'° and Rl' is from .3 to 6;
R'2, R13, R'4 and Rls are independently selected from the group consisting of C1_3-alkyl, wherein the sum of the carbon atoms in R'2, R13, R'4 and R15 is from 4 to 6.
[000102] The ring portion, L, is selected from the group consisting of:

Y~
22 2~ ~ ~ Y2 -R R

R2a ~N N
R2 -~'' \ "'S02NH2 S
N N
~2sR25 \S "'SO2NH2 and Ya N
R28R2~ ~ S02NH2 S
Y1 and YZ are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and CI_3-alkyl, wherein:
i. the C~_3-alkyl is optionally substituted witlh a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Ya is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C,_3-alkyl, wherein:
ii. the C1_3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R2' and R2' are independently selected from the group consisting of a bond and C 1_2-alkyl;
R22, Rzb and R28 are independently selected from the group consisting of NH-, -O-, and -S-;
R23 is selected from the group consisting of N=, -NHCHf=, -NHCH2CH=, -NH(CHZ)ZCH=, -OCH=, -OCHZCH=, -O(CHz)zCH=, -SCH=, -SCHzCH=, and -S(CH2)ZCH=;
RZa is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C ~ _3-alkyl, wherein:
iii. the C~_3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C,_3-alkyl an<i -(CHZ)pC(O)NH-, wherein p is from zero to 2.

[000103] A subset of these preferred mono- and bi~~-substituted compounds have the structures shown below. Such preferred compounds have a tail portion, Z, selected from the group consisting of O O
II~N~II
H
O O
~I,,~N~~) HOOC
O
R, (I~N N I) H H \C
O
II~N/-~N~,,~ II
~C
COOH
HOOC
O
R, II~ ~
\N N N~C
~COOH
HOOC HOOC

O
~ I ~ /~ /~ ~
N N N/ 'COON
~COOH
HOOC CEO
O
~~N O O N
'--COON
HOOC ~ ~d O O
I I ~ ~--. ,-HO
[000104] The sulfonamide-ring structure, L, of thesE; preferred compounds is selected from the group consisting of N
H

N ~ S02NH2 N

HNHN

NHCH

N
H
where X can be F, CI, Br, or I;
H
N
CI
CI
N
H
N N
H ~S '~S02NH2 SO?NHS

H3C\
N N
N \ ~S02NH2 S
N N
HNCH2CH2CONH \S 'S02NH2 N

N \ S
H
N

O ~ S
N

S

\ S02NH2 OCH2 , axed [000105] A subset of the above groups of compounds contains all of the compounds except those constructed with and containing the tail. piece corresponding to diethylenetriaminopentaacetic acid.
[000106] The invention also includes methods of synthesis for the described compounds. Among the mono-substituted compounds, certain compounds can be present in different isomeric forms. Thus, in one embodiment, such isorr~eric compounds are present in substantially isomerically pure form. The compounds are prepared by reacting in the presence of water and a coupling agent, a tail portion precursor in which all but one reactive carboxyl group is blocked, and a ring portion having at least one site available for reacting, preferably an amine or a hydroxyl group. The reaction occurs by mixing a tail portion and a ring portion in the presence of a coupling agent. The coupling agent is preferably a carbodiimide. More preferably, the coupling agent is a carbodiimide selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDAC) and diisopropyl carbodiimide (DCC). Following the coupling, blocking groups, (if 9any) are removed by acid hydrolysis.
[000107] The invention also includes methods of treating or preventing diseases and disorders involving elevated carbonic anhydrase activity, preferably diseases and disorders of the eye such as glaucoma and ocular hypertension. The method comprises ocular administration of the compound to the subject. The compound is administered in an amount effective in treating or preventing ocular hypertension and/or glaucoma.
SULFONAMIDE REAGENTS WITH RING PORTIONS AND REACTIVE
GROUPS

[000108] As shown in FIGS. 2A and 2B, all of the sulfonamide compounds tested to date contained a sulfonamide group (-SOzNH2) that was bonded to a carbon atom in a ring structure. Most of those compounds (including A 'through L, S, and T) used benzene rings, but several used heterocyclic rings, including thiadiazole and thiadiazoline rings (compounds M through O) and benzothiazole double-ring structures (compounds P through R). Most of these compounds were purchased from Sigma-Aldrich (St. Louis, MO) or Acros Organics (2440 Geel, Belgium). Those that could not be purchased were prepared as described in Example 1, below.

[000109) Numerous other unsaturated heterocyclic ring structures are known. More than a dozen "standard" unsaturated heterocyclic ring structures are listed in nearly any college textbook on organic chemistry, and include pyran, pyrrole, pyrrolidine, pyridine, pyrimidine, thiazole, thiophene, thiolane, i~midazole, oxazole, isoxazole, and furan. Those are just some of the single-ring unsaturated heterocyclic structures; an even greater variety of double-ring and triple-ring lheterocyclic structures are also known. In addition to those "standard" ring structures, numerous other variants are also known to skilled synthetic chemists.

[000110] The assumption that unsaturated rings are likely to perform better than saturated rings is based on prior tests of numerous sulfonamide reagents.
However, other candidate reagents containing sulfonamide groups bonded to heterocyclic rings which are saturated (such as piperidine, morpholine, pyrrolidine, thiolane, thiane, oxane, and oxolane) can be made and bonded to AAC reagents as described herein.

[000111] Any such heterocyclic ring structure which has a sulfonamide group bonded to it (or which can be modified by coupling a sulfonamide group to it), and which has (or can be given) at least one reactive group elsewhere., for reaction with a carboxyl group on an AAC reagent as shown in FIG. 1, can be evaluated for use as disclosed herein, using no more than routine experimentation.

[000112] When evaluating potential heterocyclic caazdidates for synthesis and testing, heterocyclic ring structures that already have been shown to be effective, in other CA inhibitor drugs that were sufficiently potent to merit commercialization, are likely to deserve relatively early evaluation, as preferred candidates for testing.

[000113] In the compounds which used a sulfonamide group bonded to a benzene ring, the benzene ring also had a primary amine (-NHZ) or a short-chain alcohol/hydroxy group (-CH20H or -CH2CHZOH) bonded to it. 'Chew amine or alcohol groups provided a hydrogen atom that was displaced when the reagent reacted with a carboxyl group on an AAC reagent. In FIG. 3, compounds 1 A and 6M illustrate the type of bond that will be created when an amine group is the reactive group, while compound 4S illustrates the type of bond that will be created when a hydro~:y group is the reactive group. Sulfonamide reagents containing various other types of pc,ndant reactive groups can be used, so long as the reactive group will form stable bonds when reacted with a carboxylic group in an AAC reagent.

[000114] In general, any testing of compounds that are structurally similar to the compounds that have already been tested should be planned and earned out with due regard for the enzyme inhibition data that are listed in Table 1. In particular, consideration and ranking of potential candidates for possible evaluation should bear in mind that CA inhibiting potency generally correlated with the length of the sulfonamide reagent (i.e., the number of atoms in the shortest continuous pathway between the sulfonamide group and the reactive group), and/or with the number of additional electronegative moieties attached to the ring structure. The longest sulfonamide reagents that were tested (which included reagents O through R) generally provided more potent CA inhibitors, while the shortest sulfonamide reagents that were tested (which included reagents A through C) provided weaker CA inhibitors, especially if no halogen groups were also present. Middle-length sulfonamide reagents, and short reagents with one or more halogen groups, provided intermediate CA inhibitors.

[000115] Unsaturated ring structures having a sulfonamide group bonded to one carbon atom on the ring structure, and an amino acid residue or derivative bonded to another carbon atom on the ring structure are also believed to be effective compounds [000116] Additional or alternate substituents can also be added to the unsaturated rings; including halogen atoms, lower alkyl groups, for example, C1_6 which, preferably are not too bulky. Tertiary-butyl substituents are bulkyr and less preferred.

[000117] It also should be recognized that various rr~etallo-complexes were made, by reacting transition metals, such as zinc or copper, with various AAC-sulfonamide compounds. In general, the complexes containing zinc or copper were more potent, as CA inhibitors, than the same compounds without zinc or copper.
Since zinc has no reduction-oxidation potential and is far more prevalent and widespread in the human body than copper, it is generally presumed that zinc complexes a.re likely to be preferable to metallo-complexes made with copper or other metals.
SCREENING TESTS USING ENZYME INHIBITION

[000118] When each of the seven AAC reagents shown in FIG. 1 were bonded to each of the 20 sulfonamide compounds shown in FIG. 2, a total of 140 different mono-sulfonamide compounds (and a substantial number of bis-sulfonamide compounds) were created. All of these compounds were screened in in vitro enzyme inhibition tests, to determine how potent they were in inhibiting each of three different CA isozymes (hCA-I and hCA-II expressed by cloned human genes in E. coli cells, and bovine CA-IV obtained from lung microsomes).

[000119] Enzyme inhibition tests evaluated the abiliity of various successive dilutions of each test compound to inhibit the hydrolysis of 4-nitrophenylacetate, using nM hCA-I, 3.3 nM hCA-II, and 34 nM bCA-IV. Duplicate te;>ts were done for each compound at each concentration, and all values reported were the mean of the results.
Optical density measurements were made by an automated spectrophotometer, at nanometer wavelength.

[000120] The data points listed in Table 1 represent the nanomolar concentration of a particular compound, which was effective in reducing, by 50%, the activity of a fixed quantity of hCA-I, hCA-II, or bCA-IV. Because of how these tests were done, a low value indicates that a certain compound was potent and effective as a CA inhibitor; by contrast, a high value indicates that a compound was weak as an inhibitor.

[000121] For comparative purposes, six prior known CA inhibitors were also tested against the same three CA isozymes. The prior art CA inhibitors were acetazolamide (AZA), methazolamide (MZA), ethoxzolamide (EZA), dichlorophenamide (DCP), dorzolamide (DZA), and brinzolamide (BRZ).

[000122] One of the goals of this set of in vitro enzyme screening tests was to identify either or both of the following: (i) compounds that could exceed and surpass the CA-inhibiting potencies of dorzolamide and/or brinzolamide; andlor, (ii) compounds that, unlike dorzolamide and/or brinzolamide, would be water-scduble, and that would have CA inhibiting potencies that were at least in the same ballpark as dorzolamide and/or brinzolamide.

[000123] As can be seen from the data in Table 1, a substantial number of the newly synthesized AAC-sulfonamides met and surpassed the "benchmark"
standards set by dorzolamide and brinzolamide. Accordingly, several of those compounds were selected for additional tests, as described below.

[000124] Compounds of the present invention, prefE;rably, have potent CA-II
and CA-IV activity. This is because CA-II and CA-IV are regarded as the principal CA
isozymes inside the eye. On the other hand, CA-I activity can also be important in that compounds having such activity are believed to produce minimal side effects should they permeate outside of the eyeball and enter the bloodstream. Such compounds are less likely to cause adverse should they enter the bloodstream, if they also bind fairly tightly to CA-I. CA-I is a slow-acting CA isozyme which is present at concentrations of roughly 150 ~.M. This type of binding will render them less likely to interfere with CA-II in the blood, which is a fast-acting enzyme present in blood at only about 20 ~.M. As an illustration of that principal, it is suspected that one of the reasons dorzolamide causes various undesired side effects is that it has almost no affinity for CA-I;
therefore, any dorzolamide which permeates out of the eye and enters the blood will have a much more powerful effect on CA-II enzymes in the blood.
Table 1.
Inhibition Data for AAC-Sulfonamide Compounds.
No. or Inhibitor KI (nM;~
hCA Ia hCA IIa bCA IVb Acetazolamide 900 12 220 Methazolamide 780 14 240 Ethoxzolamide 25 8 13 Dichlorophenamide1200 38 380 Dorzolamide >50,000 9 43 Brinzolamide - 3 45 1A 27000 (45400) 280 (295) 1240 (1310) 1B 23000 (25000) 233 (240) 1500 (2200) 1C 21300 (28000) 214 (300) 450 (3000) 1D 32700 (78500) 313 (320) 980 (3200) 1E 3600 (25000) 125 (170) 279 (2800) 1F 3450 (21000) 90 ( i1 257 (2500) 60) 1G 1300 (8300) 54 (fi0) 145 (180) 1H 975 (9800) 80 (1l 10) 165 (320) 1I 610 (6500) 38 (~t0) 57 (66) 1J 515 (6000) 55 (70) 114 (125) 1K 405 (6100) 24 (:!8) 107 (175) 1L 400 (8400) 69 (',~5) 145 (160) 1M 415 (8600) 50 (fi0) 265 (540) 1N 710 (9300) 13 (1.9) 120 (355) 10 280 (455) 1.5 (3) 48 (125) 1P 65 (70) 6 (9) 14 (19) 1Q 50 (55) 5 (8) 14 (17) 1R 46 (50) 6 (7) 12 (15) 1S 1260 (24000) 110 x;125) 350 (560) 1T 1200 (18000) 97 (1.10) 340 (450) 2A 25800 (45400) 280 x;295) 1200 (1310) 2B 23000 (25000) 234 (240) 1520 (2200) 2C 20600 (28000) 2101;300) 450 (3000) 2D 30500 (78500) 3181;320) 930 (3200) 2E 3500 (25000) 101 1,170) 256 (2800) 2F 3300 (21000) 88 (160) 240 (2500) 2G 1000 (8300) 53 (fi0) 136 (180) 2H 960 (9800) 82 (110) 150 (320) 2I 620 (6500) 36 (40) 53 (66) 2J 510 (6000) 57 ( 7 0) 110 (125) 2K 345 (6100) 26 (28) 100 (175) 2L 300 (8400) 66 ( 7 5) 148 ( 160) 2M 410 (8600) 52 (Ei0) 240 (540) 2N 520 (9300) 14 (l9) 106 (355) 20 268 (455) 2 (3) 45 (125) 2P 62 (70) 7 (9) 13 (19) 2Q 48 (55) 6 (8) 13 (17) 2R 44 (50) 5 (7) 10 (15) 2S 1200 (24000) 103 1;125) 350 (560) 2T 1100 (18000) 94 (110) 320 (450) 3A 20500 (45400) 286 (295) 1020 (1310) 3B 13000 (25000) 235 1240) 1005 (2200) 3C 12000 (28000) 2001;300) 365 (3000) 3D 21800 (78500) 315 1;320) 950 (3200) 3E 1760 (25000) 81 (170) 245 (2800) 3F 1540 (21000) 80 ( 160) 250 (2500) 3G 780 (8300) 49 (fi0) 103 (180) 3H 900 (9800) 78 (l 10) 155 (320) 3I 380 (6500) 35 (CEO) 54 (66) 3J 430 (6000} 50 ('.~0) 115 (125) 3K 290 (6100) 24 (:?8) 110 (175) 3L 250 (8400) 68 (75) 135 (160) 3M 250 (8600) 53 (fi0) 130 (540) 3N 275 (9300) 12 (l.9) 76 (355) 30 240 (455) 2 (3) 39 (125) 3P 56 (70) 7 (9) 12 (19) 3Q 45 (55) 5 (8) 11 (17) 3R 43 (50) 4 (7) 9 (15) 3S 1020 (24000) 106 x;125) 350 (560) 3T 1000 (18000) 95 (1.10) 330 (450) 4A 9200 (45400) 2801;295) 800 (1310) 4B 10000 (25000) 230 (240) 750 (2200) 4C 6300 (28000) 1241;300) 330 (3000) 4D 16000 (78500) 310 ( 320) 750 (3200) 4E 900 (25000) 52 (170) 83 (2800) 4F 650 (21000) 47 (160) 75 (2500) 4G 360 (8300) 31 (fi0) 40 (180) 4H 700 (9800) 52 (110) 69 (320) 4I 130 (6500) 25 (~i0) 50 (66) 4J 123 (6000) 36 ( 7 0) 78 ( 125) 4K 130 (6100) 19 (2 8) 96 (175) 4L 210 (8400) 73 ( ~ 5) 105 (160) 4M 225 (8600) 12 (Ei0) 37 (540) 4N 215 (9300) 9 (19) 26 (355) 40 200 (455) 2 (3) 23 (125) 4P 48 (70) 3 (9) 11 (19) 4Q 43 (55) 2 (8) 10 (17) 4R 39 (50) 1.5 (7) 8 (15) 4S 850 (24000) 1041;125) 335 (560) 4T 640 (18000) 89 (110) 310 (450) 4Aa 8100 (45400) 2301;295) 650 (1310) 4B2 7650 (25000) 1901;240) 620 (2200) 4C2 5000 (28000) 87 (?.00) 230 (3000) 4D2 12500 (78500) 3001;320) 510 (3200) 4E2 510 (25000) 36 (170) 62 (2800) 4F2 400 (21000) 33 (160) 54 (2500) 4G2 240 (8300) 20 (60) 35 (180) 4H2 225 (9800) 31 (110) 50 (320) 4I2 105 (6500) 15 (4~0) 34 (66) 4J2 89 (6000) 23 (70) 45 (125) 4K2 65 (6100) 13 (2.8) 51 (175) 4L2 72 (8400) 24 (75) 69 (160) 4M2 80 (8600) 6 (60) 12 (540) 4N2 105 (9300) 7 (1!)) 17 (355) 402 125 (455) 1 (3) 10 (125) 4P2 33 (70) 2 (9) 9 (19) 4Q2 37 (55) 1.5 (8) 8 (17) 4R2 25 (50) 1 (7j 6 (15) 452 460 (24000) 78 (125) 200 (560) 4T2 450 (18000) 71 (110) 165 (450) Zn-4C2 450 20 210 Zn-4E2 45 6 10 Zn-4F2 39 8 13 Zn-4M2 40 0.8 5 Zn-4N2 40 0.7 8 Cu-4M2 39 0.6 6 Al-4M2 51 0.9 8 SA 8600 (45400) 210 (295) 550 (1310) 5B 7200 (25000) 180 (240) 500 (2200) SC 4100 (28000) 75 (:400) 180 (3000) 5D 12000 (78500) 250 {320) 630 (3200) SE 550 (25000) 23 (170) 40 (2800) 5F 290 (21000) 15 (1160) 36 (2500) 5G 180 (8300) 13 (ti0) 29 (180) SH 200 (9800) 20 (l~ 10) 32 (320) SI 96 (6500) 12 (40) 27 (66) SJ 79 (6000) 16 (70) 29 (125) SK 58 (6100) 10 (:?8) 36 (175) 5L 87 (8400) 12 (';~5) 52 (160) 5M 55 (8600) 0.9 (60) 28 (540) 5N 62 (9300) 2 (19) 19 (355) 50 110 (455) 1 (3) 16 (125) 5P 36 (70) 0.8 (9) 9 (19) SQ 32 (55) 0.8 (8) 6 (17) 5R 30 (50) 0.6 (7) 5 (15) SS 510 (24000) 80 (125) 265 (560) ST 360 ( 18000) 61 ( 1.10) 190 (450) SA2 5400 (45400) 175 x;295) 330 (1310) SB2 6400 (25000) 155 (240) 425 (2200) 5C2 3750 (28000) 64 (300) 156 (3000) SDZ 9800 (78500) 215 1;320) 430 (3200) SEZ 380 (25000) 19 (170) 37 (2800) SFZ 170 (21000) 8 ( 1 Ei0) 25 (2500) SGZ 135 (8300) 7 (6(I) 18 (180) SHZ 155 (9800) 15 (110) 27 (320) SI2 79 (6500) 10 (40) 21 (66) SJZ 70 (6000) 9 (7(I) 24 (125) SK2 43 (6100) 9 (2f.) 25 (175) 5L2 63 (8400) 11 ('75) 41 (160) SMZ 50 (8600) 1 (60) 7 (540) SN2 54 (9300) 1.5 (19) 9 (355) SOa 102 (455) 0.6 (3) 8 (125) 5P2 25 (70) 0.5 (9) 6 (19) 5Q2 21 (55) 0.6 (8) 5 (17) SRZ 16 (50) 0.5 (7) 4 (15) SSa 325 (24000) 43 (1! 25) 170 (560) STZ 290 (18000) 34 (1.10) 115 (450) Zn-5C2 350 16 100 Zn-5E2 39 9 21 Zn-5F2 36 7 16 Zn-SMZ 40 0.5 4 Zn-5N2 43 0.4 5 Cu-5M2 31 0.3 3 AI-5M2 44 0.8 6 6A 8900 (45400) 200 x;295) 545 (1310) 6B 7450 (25000) 195 ( 240) 500 (2200) 6C 4000 (28000) 79 (300) 170 (3000) 6D 12500 (78500) 2651,320) 620 (3200) 6E 610 (25000) 30 (170) 48 (2800) 6F 290 (21000) 18 (160) 32 (2500) 6G 210 (8300) 17 (Ei0) 31 ( 180) 6H 230 (9800) 16 (l 10) 30 (320) 6I 78 (6500) 10 (~~0) 32 (66) 6J 76 (6000) 13 ( 7 0) 24 (125) 6K 50 (6100) 12 (a8) 35 (175) 6L 93 (8400) 16 ( 7 5) 65 (160) 6M 54 (8600) 2 (6U) 21 (540) 6N 66 (9300) 2 (1ST) 16 (355) 60 120 (455) 1.5 (:3) 13 (125) 6P 39 (70) 1 (9) 7 (19) 6Q 33 (55) 0.9 (.B) 5 (17) 6R 30 (SO) 0.8 ('7) 6 (15) 6S 500 (24000) 72 (125) 250 (560) 6T 430 (18000) 56 (110) 155 (450) 6A2 5600 (45400) 160 (;295) 320 (1310) 6B2 6150 (25000) 145 ( 240) 400 (2200) 6C2 3800 (28000) 60 (300) 150 (3000) 6D2 9500 (78500) 210 (;320) 400 (3200) 6E2 350 (25000) 15 (170) 39 (2800) 6F2 160 (21000) 7 (160) 20 (2500) 6G2 155 (8300) 7 (6C~) 16 (180) 6H2 150 (9800) 9 (110) 22 (320) 6I2 85 (6500) 12 (4.0) 25 (66) 6J2 76 (6000) 8 (70) 20 (125) 6K2 49 (6100) 10 (28) 21 (175) 6L2 50 (8400) 13 (',~5) 36 (160) 6M2 55 (8600) 2 (60) 4 (540) 6N2 36 (9300) 1.1 (19) 5 (355) 60Z 76 (455) 0.5 (3) 6 (125) 6P2 23 (70) 0.5 (9) 4 ( 19) 6Q2 21 (55) 0.6 (8) 5 (17) 6R2 15 (50) 0.6 (7) 5 (15) 6S2 335 (24000) 38 (1.25) 150 (560) 6T2 300 (18000) 33 (110) 96 (450) Zn-6C2 300 11 76 Zn-6E2 35 9 16 Zn-6F2 30 6 12 Zn-6M2 27 0.7 8 Zn-6N2 32 0.8 7 Cu-6M2 21 0.2 2 Al-6M2 19 0.7 3 7A 12500 (45400) 2801;295) 800 (1310) 7B 10000 (25000) 235 1;240) 860 (2200) 7C 11500 (28000) 175 1;300) 320 (3000) 7D 21000 (78500) 3101;320) 840 (3200) 7E 1200 (25000) 69 (170) 135 (2800) 7F 1050 (21000) 61 (160) 215 (2500) 7G 450 (8300) 43 (E~0) 79 (180) 7H 870 (9800) 67 ( 110) 108 (320) 7I 175 (6500) 34 (4~0) 56 (66) 7J 220 (6000) 47 (70) 90 (125) 7K 250 (6100) 23 (2;8) 105 (175) 7L 240 (8400) 65 ( 7 5) 123 ( 160) 7M 245 (8600) 54 (E0) 105 (540) 7N 250 (9300) 13 (19) 57 (355) 70 210 (455) 3 (3) 35 (125) 7P 52 (70) 6 (9) 13 ( 19) 7Q 47 (55) 4 (8) 12 (17) 7R 41 (50) 3 (7) 7 (15) 7S 945 (24000) 110 ( 125) 320 (560) 7T 760 (18000) 94 (110) 300 (450) a Human (cloned) isozymes; b From bovine lung microsomes, by the esterase method.

SOLUBILITY, PARTITION, AND TISSUE PERMEATION TESTS

[000125] Several additional in vitro tests were also run on a number of promising AAC-sulfonamide compounds. Those tests included water solubility (using an aqueous buffer, pH 7.4, at 25°C), and two-phase partitioning (us:ing aqueous buffer, and chloroform). The data are in Table 2. In the two-phase partitioning tests, the number in Table 2 represents the log (base 10) of the ratio between the concentration of the test compound in the organic solvent, divided by its concentration in aqueous solvent;
accordingly, a lower number represents greater solubility in water. In comparing the numbers for the test compounds against the control number (2.0) for dorzolamide, it should be kept in mind that (i) the dorzolamide solution was much more acidic than the test compounds (all of which were tested at pH 7.4, which is slightly basic, and which presumably would have shown significantly higher values if tested at pH 6.5 or 6.8); and, (ii) because a logarithmic scale was used, even a slightly lower number indicates substantially higher water solubility.

[000126] In vitro tissue permeation tests were also performed, using corneal tissue from rabbits, both with and without the epithelial layer. A;; described in Example 10, these tests used a segment of rabbit corneal tissue, which wa;> placed in a window between two fluid chambers, one of which initially contained a known concentration of the test compound. These tests were intended to evaluate the ability of candidate compounds to permeate through the corneal tissue and into the aqueous and vitreous humors (liquids) inside the eye, after topical application in the form of eyedrops.

[000127] The results of these tests are presented in 'Cable 2. These data clearly indicate that the AAC-sulfonamide compounds that were tested performed quite well in these water solubility and tissue permeation tests, compwed to dorzolamide.

Aqueous Solubility, Aqueous Vs. Organic Solvent Partitioning, and In Vitro Corneal Tissue Permeation Compound pH Solubility,Log, k;" x 103 (hru) mMol org/aq. Cornea intactNo epithelium Dorzolamide 5.5 60 2.0 3.0 5.2 7.4 62 1.230 2.5 4.6 2R 7.4 37 1.925 5.1 10.3 3Q 7.4 44 1.166 4.7 8.4 4N 7.4 65 1.750 4.8 9.5 4N2 7.4 41 1.333 3.1 5.5 4K2 7.4 32 1.764 3.6 7.0 402 7.4 46 1.150 3.0 5.9 SC 7.4 48 0.983 3.8 6.9 5C2 7.4 35 1.456 3.6 6.5 5M 7.4 59 1.375 4.3 8.1 5M2 7.4 54 1.961 3.9 6.2 6I 7.4 50 1.747 3.3 6.5 6M 7.4 52 1.446 4.6 8.4 6GZ 7.4 47 1.534 4.5 8.3 70 7.4 44 2.354 5.1 10.7 [00012$] In addition to the in vitro permeation tests described above, in vivo permeation tests also were carried out on two test compounds, to measure how much of the test compounds were found in various ocular tissues and fluids after administration of eyedrops. These tests are described in Example 11, and the resuha are provided in Table 6.
[000129] In order to provide a consistent standard for evaluating and comparing solubility of various compounds, pH 6.5 has been selected and referred to in the claims. It is slightly acidic, to a point of beginning to raise questions about potential irritation in highly sensitive patients, but not to a point of posing a severe threat of eye irritation in most patients.
[000130] It should be noted that many of the AAC-sulfonamide compounds disclosed herein have sufficient solubility in neutral solutions (pEi 7.0) to allow them to be formulated in aqueous eyedrops that have neutral or near-neutral pH
(regarded herein as pH levels between about 6.8 and about 7.2). In highly sensitive patients, neutral or near-neutral solutions (or solutions that are slightly alkaline, with a pH up to about 7.5, since many physiological fluids have pH levels of about 7.4) pose less risk of irntating eyes than solutions having pH levels below about 6.8, especially when it comes to eyedrops that will be applied on a repeated and chronic basis, two or more times each day, for the rest of a patient's life. Therefore, AAC-sulfonamide ~~ompounds that have sufficient levels of water solubility to create 2% w/w aqueous solutions with a pH
anywhere between 6.8 and 7.2 are regarded as the most highly preferred candidates for evaluation for use as disclosed herein.
[000131] In addition, a somewhat broader pH range which covers the ophthalmic "comfort range" as discussed above extends from pHI 6.6 up to pH
7.8, is regarded as being substantially preferable to existing dorzolamide eyedrops, which have a pH of about 5.6, and brinzolamide eyedrops, which have a pH of 7.5, but which must be administered as a suspension rather than a solution. Clearly, eyedrops having a pH of down to about 6.3 will still run a substantially lower risk of causing burning and itching than eyedrops having a pH of 5.6. Accordingly, when compared to the existing CA
inhibitor eyedrops that are currently available to the public, candidate AAC-sulfonamide compounds as disclosed herein which cannot reach the benchmark 2% w/v solubility levels in water, except in a pH range below 6.8 and down to abo~zt 6.3, may contain various candidate compounds that would merit testing in animal or human trials.
Nevertheless, since excellent results have been obtained to date by compounds that can indeed meet the desired solubility levels in neutral pH ranges, those compounds are regarded as preferred compounds.

TESTS ON NORMOTENSIVE RABBITS
[000132] The first set of in vivo animal tests used "normotensive" rabbits.
These are rabbits which have normal IOP levels; by contrast, "glaucomatous"
rabbits are described below.
[000133] In all IOP tests on either nor~rnotensive or glaucomatous rabbits, the drug was administered to only one eye, leaving the other eye as an untreated control.
To minimize any diurnal, seasonal, inter-individual, or other var-i ations, the reduction in IOP caused by a test drug was calculated by subtracting the IOP in the treated eye from the IOP in the untreated eye. To further reduce the possibility of inaccurate measurements, IOP was measured three times for each eye at ea<;h time interval; the mean value of those three measurements was recorded and used for all subsequent calculations. In addition, the pneumatonometer was checked at lf;ast twice each day, using a calibration verifier. A drop of an ocular anesthetic (oxybuprocaine HCI, diluted 1:1 with saline) was placed in the eye immediately before each set of pressure measurements.
[000134] The tests on normotensive rabbits were carried out using a number of AAC-sulfonamide compounds that were selected based on the results of the enzyme inhibition tests described above. These tests also used dorzolamide as a control, for comparative purposes.
[000135] The results are provided in Table 3. As indicated therein, a number of the compounds tested provided substantially better and more potent IOP-lowering results than dorzolamide, at all time intervals tested (i.e., 30 minutes, 60 minutes, and 90 minutes after the test drug was administered).

IOP Reductions in Normotensive (23.0 ~ 1.5 mm~ Hg) Rabbits DIOP (mm Hg) Inhibitor pH t = 30 min t = 60 min t = 90 min Dorzolamide 5.5 1.9 0.2 4.0 0.3 2.1 t 0.2 Brinzolamide**5.5 2.9 t 0.1 3.2 0.3 6.3 0.4 7.0 2.2 0.2 4.5 0.3 5.7 ~ 0.3 2R 7.0 2.60.2 4.9f0.3 6.210.3 4K 7.0 1.8f0.2 2.50.3 3.410.3 4N 7.4 6.0 0.1 9.3 0.2 1 2.2 0.4 Zn-4N susp. 6.5 4.9 0.1 9.5 0.2 13.1 t 0.5 5C 7.0 3.1 t 0.2 4.5 0.2 7.2 t 0.3 5C2 7.0 4.4 t 0.2 7.5 t 0.2 9.5 t 0.3 5M 7.4 3.1 0.5 6.8 0.2 4.5 t 0.3 5M2 7.5 6.5 0.1 13.0 t 0.2 8.1 0.6 Zn-5M2 susp. 7.0 7.4 t 0.2 14.5 0.2 9.5 t 0.3 5N 7.5 6.7 t 0.1 9.2 0.3 13.0 0.4 6I 7.0 1.2 t 0.1 3.9 0.15 7.2 t 0.2 Zn-6I 6.2 2.9 0.1 5.6 0.1 8.6 t 0.2 6M 7.0 4.6 0.2 8.9 0.4 12.1 0.4 Zn-6M** 6.0 5.20.1 10.30.3 13.610.5 Cu-6M* * 6.0 4.7 t 0.2 9.6 0.2 13.5 t 0.3 70 7.0 2.5 0.3 4.9 0.3 8.0 f 0.4 [000136] In addition, another set of in vivo tests waG done on normotensive rabbits, to further evaluate the duration of relief (i.e., the reduction in IOP levels as a function of time, over several hours) that could be provided selected AAC-sulfonamide compounds. Those tests compared 2% solutions of compounds ~N (in both trisodium salt form and zinc complex form, both at pH 7.0) and SMZ (trisodium salt and zinc complex, both at pH 7.0) against dorzolamide and brinzolamide (both as HCI salts, pH
5.5). The results are presented graphically in Figure 4. As shown therein, both AAC-sulfonamide compounds that were tested outperformed both dorzolamide and brinzolamide, in terms of (i) magnitude of peak relief, (ii) duration of substantial reductions in IOP, and (iii) total "area-under-the-curve" IOP reductions, which offer an indi~~ator of total cumulative benefit from a single dosage.
[000137] As a benchmark for evaluation and comparison purposes, a fixed time of 4 hours after the eyedrops are administered, is regarded ass a useful standard time for measuring long-lasting IOP reduction. This standard is applied by measuring IOP
reductions in normotensive rabbit eyes, which provide more consistent results than diseased and glaucomatous eyes, which suffer much higher levels of variability when measured. At the 4-hour mark, a 3 mm Hg reduction in IOP, when tested on normotensive rabbit eyes, has been chosen as a suitable "benchmark" level for measuring long-lasting IOP reductions. Neither dorzolamide nor brinzolamide eyedrops are believed to be able to meet the 3 mm Hg IOP reduction level, at 4 hours after administration. By contrast, several AAC-sulfonamide compounds tested to date have easily surpassed that standard.
TESTS ON GLAUCOMATOUS RABBITS
[000I38] The second set of in vivo animal tests used "glaucomatous"
rabbits. These are rabbits which have abnormally high IOP level:>. This condition was created by surgically injecting a-chymotrypsin into one eye, as described in Melena et al 1999, then waiting at least a month. The chymotrypsin enzyme mildly damaged certain structures in the eye, in a manner which hindered but did not destroy the eye's ability to properly drain the aqueous humor.
[000139) Because of the additional costs and burder.~s of tests on glaucomatous rabbits, only a few AAC-sulfonamide compounds were tested on them; the selected compounds were chosen, based on their performance with normotensive rabbits.

Dorzolamide was also selected, for comparative purposes.
[000140] The results are in Table 4. As shown therein, all of the AAC-sulfonamide compounds that were chosen and tested outperformed dorzolamide by wide margins, in terms of both (i) the extent of peak relief provided, and (ii) the lasting duration of relief provided.
[000141] In Table 4, it also should be noted that the pH of all three AAC-sulfonamide formulations were much closer to neutral than the dorzolamide formulation, which was acidic (pH 5.6). Therefore, the AAC-sulfonamide formulations posed a much lower risk of eye irritation than the acidic dorzolamide formulation.

IOP Reductions in Glaucomatous (34.5 ~ 2.0 mm Hg) Rabbits DIOP (mm Hg)*

Inhibitor pH t = 30 t = 60 min t = 90 min min Dorzolamide 5.6 3.6 0.2 6.7 0.3 4.2 0.2 4N 7.4 7.3 0.1 11.3 0.4 1 5.2 0.4 SCZ 7.0 6.00.4 12.50.2 11.00.2 5M2 7.5 7.4 0.3 13.0 0.5 14.9 0.5 5N 7.5 8.8 t 0.6 16.0 0.4 17.5 0.3 Zn-5Ma susp 7.0 19.8 0.7 19.2 0.8 18.9 0.5 6M 7.0 7.6 0.4 12.910.5 15.1 10.5 [000142] By lowering intraocular pressures, and thereby helping to ensure and promote proper levels of blood flow through the capillaries in the retina and other ocular tissues, the compounds disclosed herein can help treat not just glaucoma, but other eye diseases as well. As examples, it recently has been reported that certain combinations of high-dosage anti-oxidants (including Vitamin C, Vitamin E, and zinc) can help beat or retard macular degeneration (Age-Related Eye Disease Study Group, Arch.

Ophthalmology 119: 1417-1436 (2001), and that zeaxanthin, an anti-oxidant and ultraviolet-absorbing carotenoid pigment found in the macula, in the center of the retina, can also help prevent and treat macular degeneration (e.g., US 5"827,652, Garnett et al 1998). Those compounds must be deposited into various eye tissues (including retinal tissues) in order to provide their protective effects. Accordingly, the CA
inhibitor eyedrops of this invention can be used in conjunction with other therapeutic compounds, to help promote higher levels of deposition of those therapeutic compounds in retinal or other ocular tissues. As such, certain claims refer to CA inhibitor eyedrops as disclosed herein, for use in "reducing intraocular pressure". That phrase is intended to include any form of treatment, using such compounds, which reduces IOP, rc;gardless of whether a patient is suffering from ocular hypertension, glaucoma, macular degeneration or edema, or any other ocular disorder, and/or is being treated for any other condition in which increased blood flow through the macula, retina, or any other ocular tissue is desired (such as during therapy in which anti-oxidants, zeaxanthin or other carotenoids, or any other nutritional or therapeutic compound is being administered 'to the patient to treat or improve an ocular condition).
[000143] Similarly, the CA inhibitor eyedrops of this invention can be used in conjunction with various other compounds, to provide greater IOP
reductions, in patients in need of such treatment. Examples of such other compounds include prostaglandins and prostaglandin derivatives (such as latanopros~t, a prostaglandin analog sold under the trademark XALATAN by Pharmacia-Upjohn), beta-blockers (such as timolol, a beta-adrenergic receptor blocker sold under the trademark TIMOPTIC
by Merck), etc.
[000144] The compounds of this invention may also be useful for treating other diseases and disorders, which are entirely unrelated to vision. For example, as described in Supuran and Scozzafava et al 2002 and in various other articles cited therein, there are substantial reasons to suspect that CA inhibitor drugs may be effective in treating some forms of cancer and osteoporosis, and some types of microbial infections, including malaria. Accordingly, the compounds disclosed herein can be tested, to evaluate their efficacy in treating any such disease or disorder.
[000145] The compounds of the present invention acre prepared in pharmaceutically acceptable formulations for topical ocular administration to a mammalian subject. Such ophthalmic formulations comprise a carbonic anhydrase inhibitory compound or a pharmaceutically acceptable salt thereof, in an ophthalmically acceptable vehicle.
[000146] The mammalian subject is, preferably, a human although companion animals, such as dogs and cats, farm animals, exotic animals such as zoo animals and the like are included within the scope of the term mammalian subject as used herein.
[000147] The carbonic anhydrase inhibitory compounds of the present invention are present in the ophthalmic composition in amounts of, preferably, from about 0.05 % to about 5 % by weight, depending upon the carbonic anhydrase activity of the particular compound in the composition. Topical ophthalmic; administration is typically from 1 to 5 times daily with a daily dosage of from about 0.1 mg to about 50 mg.
(000148] The carbonic anhydrase inhibitory compounds are in a vehicle which is non-irntating and non-toxic to the tissue of the eye. Such vehicles are known in the art and commonly used in topical ophthalmic medications (sc;e for example pages 13-1 S of Physicians 'Desk Reference for Ophthalmology, 28th Ed., 'J~elsbecker et al., Eds., Medical Economics Company, Inc., Montvale, NJ, 2000, which is incorporated in its entirety by reference). The ophthalmic compositions are in a form suitable for topical ocular administration. The compositions are, preferably, sterile ;end of an ophthalmically acceptable pH, tonicity and viscosity.
[000149] The desired pH of the composition is achieved by incorporating buffering agents into the composition. Preferably, the vehicle is comprised of water although non-aqueous vehicles can be used such as alcohols, polyols and the like. The aqueous vehicle can contain various salts and buffering agents such as alkali metals and alkaline earth metals, carbonates, bicarbonates, borates, citrates, phosphates and the like to achieve a desired pH. Preferably, the pH of the compositions is within the ocular comfort region of from about 6.6 to about 7.8 and, more preferably, the pH is within the range of normal tears, i.e. from about 7.0 to about 7.5.
[000150] The composition also, preferably, has a toxicity within a range tolerable to the subject. Such a tolerable tonicity range is considered to be that which is equivalent to from about 0.5-1.5% and, preferably, about 0.9% mass/volume (about 300 mosM/L) sodium chloride (see, for example, Fasihi et al., S. Afr. Med. J.
75:233-235, 1989). The desired tonicity of the composition can be achieved by adjusting the amount and composition of various salts and buffering agents in the composition.
[000151] The viscosity of the composition is also within an acceptable range for tolerance by the subject, i.e. from about 1 to about 15 centipoise (Id.).
The viscosity can be adjusted by including in the composition, for example, cellulose-derived viscosity adjusting agents such as carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methylcellulose.
[000152] In addition, various other components can be present in the composition in an admixture with the aqueous solution. Such additional components can include, for example, polyvinyl alcohol, polyethylene glycol, po~ridone, dextran 70, dextrose, glycerin and the like. Various excipients which can be included in ophthalmically acceptable topical solutions are disclosed U.S. Patents Nos.
5,591,426, 5,607,698, 5,800,807 and 6,264,935, which are incorporated in their entireties by reference.
[000153] The composition can also contain, in certain embodiments, wetting agents such as Tween 80 and anti-bacterial agents such as benzalkonium chloride, parabens, chlorobutanols, thimerosal and the like.
[000154] The compounds of the present invention are, preferably, administered in eye drops, however, any mode known in the art, which is suitable for topical administration to the eye can be used.
[000155] Other routes of administration can also be used including oral administration in tablets, capsules, liquid formulations and the like as well as administration by an other routes such as intravenous, subcutaneous, intramuscular, intranasal, intraperitoneal, intrathecal, and the like. Administration of the carbonic anhydrase inhibitory compounds of the present invention by such other routes is, preferably, in an amount of from about 50 mg up to about 1500 given from 1 to 4 times daily. Dosage amounts will vary depending on such factors as the activity of the particular compound, the selected route of administration as well as factors such as subject tolerance and response.
Industrial A~nlicability [000156] The compounds disclosed herein have industrial utility, in the commercial manufacture of medicaments for treating and preventing glaucoma and other ocular and ophthalmic disorders. These compounds may also have various other types of industrial applicability as well, such as in the commercial manufacture of medicaments for treating cancer, osteoporosis, malaria, and possibly other disorders as well.
EXAMPLES
EXAMPLE l: REAGENT SOURCES AND METHODS
[000157] Amine-carboxy (AAC) reagents 1 through 7, as shown in FIG. 1, as well as any anhydrides, carbodiimides, solvents, or metal salts or other inorganic reagents, were purchased from commercial suppliers (see Table '.i).

Names and Sources of Sulfonamide Reagents Letter Source or synthesis in Sulfonamide Reagents Shown in Fig.
Fi 2 .

A 2-amino-benzenesulfonamide Si a-Aldrich B 3- amino-benzenesulfonamide See Exam 1e 1 C 4- amino-benzenesulfonamide Si a-Aldrich D 4-h drazino-benzenesulfonamide Cri a et al 1941 E 4-aminometh 1- benzenesulfonamide Si a-Aldrich F 4- 2-aminoeth 1 - benzenesulfonamide Acros G 3-fluoro-4-amino-benzenesulfonamide Cin olani 1948 H 3-chloro-4-amino-benzenesulfonamide Cin olani 1948 I 3-bromo-4-amino-benzensulfonamide Cin olani 1948 J 3-fluoro-4-amino-benzenesulfonamide Cin olani 1948 K 4,5-dichloro-6-amino-benzene-1,3-disulfonamideLederle L 6-chloro-4-amino-benzene-1,3-disulfonamideMerck M 5-amino-1,3,4-thiadiazol-2-sulfonamideSee Exam 1e 1 N 5-imino-4-methyl-2-sulfonamido-8'-1,3,4-Scozzafava et al thiadiazoline 1998 _57_ O 5-(2-aminoethylcarboxamido)-1,3,4-thiadiazol-2-Jitianu et al sulfonamide 1997;
Su uran et al P 6-amino-benzothiazol-2-sulfonamide Eller et al 1985 6-h drox -benzothiazol-2-sulfonamide Eller et al 1985 R 6- 2-h drox eth lox -benzothiazol-2-sulfonamideEller et al 1985 S 4-h drox meth I-benzenesulfonamide Cin olani 1948 T 4- 2-h drox eth 1 -benzenesulfonamide Cin olani 1948 [000158] All the sulfonamides purchased from commercial suppliers were the highest grade or purity available and were used without additional purification. Those sulfonamides which were not purchased were prepared as follows. Compound B was prepared by hydrogenation (using hydrazine and palladium) of 3~-nitrobenzene-sulfonamide (purchased from Sigma-Aldrich). Compound D way, prepared by diazotization of sulfanilamide, followed by reduction of the diazonium salt with tin(II) chloride, as described in Crippa et al 1941. Halogenated compounds G through J
were prepared by halogenation of sulfanilamide (purchased from Sigma-Aldrich), as described in Cingolani 1948. Compound M was prepared by deacetylation of acetazolamide (purchased from Sigma Aldrich), with concentrated HCI. Compound N was prepared by deprotecting methazolamide with concentrated HCI, by the method of Jitianu et al 1997.
Compound O was prepared by acylating 5-amino-1,3,4-thiadiazole-2-sulfonamide (obtained from acetazolamide, by the method of Jitianu et al 1997) with the phthalimido derivative of beta-alanine, followed by hydrazinolysis, by the method of Barboiu et al 1999. Compounds P, Q, and R were obtained by the methods described in Eller et al 1985 and Woltersdorf et al 1989. Compounds S and T were prepared from the corresponding amines, by diazotization followed by hydrolysis of the diazonium salts, in a manner similar to compound D. These synthetic routes have also been described in other publications, including Scozzafava et al 1999a and 1999b, Scozzafava et al 2000, Casini et al 2000, and Chen et al 2000.
[000159] Acetone, acetonitrile, DMF and other solvents used in synthesis or chromatography were doubly-distilled, and kept on molecular sieves in order to maintain them in anhydrous conditions.
[000160] To protect certain types of vulnerable moieties from becoming involved in undesired reactions, tertiary-butoxycarbonyl (commonly known as "BOC") protective groups were added to the secondary amine groups in acids 1 and 3, and to the phenolic hydroxy groups in acid 7, before those particular acids were reacted with a sulfonamide reagent. The BOC protective groups were added using procedures described in Itoh et al 1980. After condensation with a sulfonamide was completed, the BOC
protective groups were removed using trifluoroacetic acid (TFA1 in methylene chloride (CH2C12).
[000161] Commercial preparations of dorzolamide ;Merck & Co., TRUSOPTR eye drops) and brinzolamide (Alcon Laboratories, AZOPTR eye drops) were used for control tests.
[000162] Melting points were recorded with a heating plate microscope and were not corrected. IR spectra were recorded in KBr pellets with a Carl Zeiss instrument. 'H-NMR spectra were recorded in DMSO-db or TFA, as solvents, using a Bruker CPX200 or Varian 300 instrument. Chemical shifts are reported as 8 values, relative to Me4Si as an internal standard. Elemental analyses were done by combustion for C, H, N with an automated Carlo Erba analyzer, and gravimetrically for the metal ions, and were generally within D 0.4% of the theoretical values, High resolution mass spectroscopy (HRMS) was recorded with an AEI-MS 902 spectrometer, using fast atom bombardment (FAB) or electrospray techniques. All reactions were monitored by thin-layer chromatography (TLC) using 0.25-mm precoated silica gel plates (E.
Merck).
Analytical and preparative HPLC was performed on a reverse-phase C,g Bondapack column, with a Beckman EM-1760 instrument.
EXAMPLE 2: PREPARATION OF MONO-SULFONAMIDE COMPOUND 4M
[000163] As an illustrative example which disclose~~ the steps used to synthesize a mono-sulfonamide compound from a sulfonamide reagent having an amine reactive group, this example provides the specific steps used to create compound 4M.
Compound 4M was synthesized by the method for synthesis of mono-sulfonamide AAC-sulfonamide compounds shown in FIG. 4. The synthesis scheme applied to compound 4M is shown in FIG. 10.
[000164] A 1 mmole quantity (180 mg, based on calculated molecular weight) of sulfonamide reagent M (shown in FIGS. 2A and 5A) was dissolved in 25 mL
of anhydrous acetone. A stoichiometric amount ( 1 mmole, based on calculated molecular weight) of AAC reagent 4 (which is EDTA) was added, with stirring. 190 mg (1 mM) of the hydrochloride salt of 1-ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide (EDAC! ! ! ) was then added, and the reaction mixture was stirred at room temperature for 15 minutes.
30 ~.L (2mM) of triethylamine was then added, to neutralize HCl from the EDAC-HCl salt, and stirring was continued at 4°C, until thin layer chromatography indicated that the desired reaction was complete; this usually required between 8 a~zd 16 hours.
[000165] The acetone solvent was then removed under a mild vacuum. The residue was suspended in ethyl acetate (5 mL), poured into a 5% solution of sodium bicarbonate (5 mL), brought to pH 7 with 1N HCI, and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and filtered, and the solvent was removed under mild vacuum.
[000166] To purify the resulting AAC-sulfonamide compound, it was dissolved in 10 to 20 mL of potassium phosphate buffer, and the solution was passed through a reverse-phase using CIg reverse-phase p-Bondapack columns, with an elution buffer containing 90% acetonitrile, 8% methanol, and 2% phosphate buffer, pH
7.4, at 10 mL/min. Impurities normally eluted within about 5 to 10 minutes, and product normally eluted after about 15 to 20 minutes.
[000167] Analysis to confirm the proper structure oi.-'the resulting AAC-sulfonamide compound 4M was carned out, resulting in the following data:
melting point: 155-6°C (dec.); IR (KBr), cm'': 1164 (SOZS''m), 1360 (502''s), 1561 (amide II), 1600 (amide I), 1765 (COOH); 3330 (NH, NH2); 'H-NMR (D20-KOD), 300 MHz, 8, ppm:
3.35 (6H, s, 3CHZCOOH); 3.54 (4H, t, CHZCHZ); 3.56 (2H, s, C~>r2CONH); the S02NH2, CONH and COOH protons are not seen in this solvent mixture, being in fast exchange, '3C-NMR (D20-KOD), 8, ppm: 53.24 (CHZCH2); 53.70 (CH2CHz); 60.18 (CHZCONH);
61.34 (CH2COOH); 159.5 (C-2 of thiadiazole); 170.3 (C-5 of thiadiazole);
176.41 (CONH); 180.82 (COOH); elemental analysis C~ZHIgN6O9S2.
EXAMPLE 3: PREPARATION OF MONO-SULFONAMIDE C'.OMPOUND 4S
[000168] The steps used to synthesize a mono-sulfonamide compound from a sulfonamide reagent having a hydroxy reactive group were essentially the same as for sulfonamide reagents having amine reactive groups, since both t~rpes of reactions used carbodiimide to remove a hydrogen proton from the sulfonamide reagent's reactive group, and a hydroxy group from the AAC reagent. The only substantive difference was that the use of a hydroxy reactive group (as appeared in sulfonamide reagents Q-T, shown in FIG. 2B) led to an ester linkage, as shown in Fig.l2, wihile the use of an amine reactive group (as appeared in sulfonamide reagents A-P, shown in FIG. 2A) led to an amide linkage, as shown in FIG. 11.
[000169] A 1 mMole of sulfonamide S was dissolved in 25 mL of anhydrous acetone. A stoichiometric amount ( 1 mmole) of AAC reagent 4 (which is EDTA) was added, with stirnng. 190 mg (1 mM) of the hydrochlloride salt of I-ethyl-3-(3-dimethyl-aminopropyl)-carbodiimide (EDChHCI) was added, and the reaction mixture was stirred at room temperature for I S minutes. 30 pL (2mM) of triethylamine was then added, to neutralize HCl from the EDChHCI salt. Stirring was continued at 4°C, until thin layer chromatography indicated that the desired reaction was connplete; this usually required between 8 and 16 hours.
[000170] The acetone solvent was then removed under a mild vacuum. The residue was suspended in ethyl acetate (5 mL), poured into a 5% solution of sodium bicarbonate (5 mL), brought to pH 7 with 1 M HCI, and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and filtered, and the solvent was removed under mild vacuum.
[000171] The resulting AAC-mono-sulfonamide compound was purified by preparative HPLC, using a CIg reverse-phase ~.-Bondapack column, with an elution buffer containing 90% acetonitrile, 8% methanol, and 2% phosphate buffer, pH
7.4, at 10 mL/min. Impurities normally eluted within about 5 to 10 minute;, and product normally eluted after about I S to 20 minutes.
[000172] Analysis to confirm the proper structure o~~the resulting AAC-sulfonamide compound 4S was carried out, resulting in the following data:
melting point: 151-2 °C; IR (KBr), cni': 1164 (SO2~'m), 1330 (S028S), 1680 (COO), 1765 (COOH); 3300 (NHZ); 'H-NMR (D20-KOD), 300 MHz, 8, ppm: 3.35 (6H, s, 3CHZCOOH); 3.54 (4H, t, CH2CHz); 3.56 (2H, s, CHZCOOCHZ); 4.49 (s, 2H, CHZ
from hydroxymethyl-benzenesulfonamide), 7.46 (d, 2H, AA~B ; 8.3), 7.78 (d, 2H, AA~B', 8.3), the SOZNH2, and COOH protons are not seen in this solvemt mixture, being in fast exchange, '3C-NMR (DZO-KOD), 8, ppm: 41.58 (CHZ, CHZ from hydroxymethyl-benzenesulfonamide), 53.24 (G'HZCH2); 53.70 (CHZCHZ); 60.18 (CHZCOO); 61.34 (CHZCOOH); 125.68 (C2/C3 -Ph), 127.41 (C3/C2 -Ph), 142.53(C1/C4 -Ph), 144.19 (C4/C 1 -Ph), 176.41 (COO-CHZ); 180.82 (COOH); Anal., found: C, 44.39; H, 5.13; N, 8.93; C1~H23N301oS requires: C, 44.25; H, 5.02; N, 9.11%.
EXAMPLE 4: PREPARATION OF MONO-SULFONAMIDE COMPOUND 1 C
[000173] AAC reagents 1, 3 and 7 were modified by the addition of protective tert-butyl-oxycarbonyl (BOC) groups to the corresponding secondary amino and hydroxy groups present in these reagents. The bulky BOC guoups displace the hydrogen in the corresponding amino (in AAC reagents 1 and 3) or hydroxyl groups (in AAC reagent 7) thus preventing sulfonamides A-T from reacting, with the BOC-protected group. Thus, a single BOC-protective group was added to the amino group of reagent 1, two BOC-protective groups were added to the two amino groups of reagent 3, and two BOC-protective group were added to the two hydroxyl groups of reagent 7.
Methods of adding BOC-protective groups are described in the literature and are well known to those skilled in the art. For example, one method of adding a BOC-protective group is by treating the corresponding AAC reagent with tert-butyl-oxycarb~onyl-azide.
[000174] The synthesis scheme for AAC-sulfonamide compound 1 C is shown on FIG. 12. To create compound 1 C, a 1 mmole quantity of sulfonamide reagent C
was dissolved in 25 mL of anhydrous acetone. A stoichiometric amount ( 1 mmole, based on calculated molecular weight) of BOC-protected AAC reagent 1 was added, with stirnng. A 1 m~mole quantity of a EDCI~HCI was then added, anal the reaction mixture was stirred at room temperature for 15 minutes. 30 ~,L (2mM) of triethylamine was then added, to neutralize HCl from the EDCI-HCl salt. Stirring was continued at 4°C until thin layer chromatography indicated that the desired reaction was connplete; this usually required between 8 and 16 hours.
[000175] The acetone solvent was then removed under a mild vacuum. The residue was suspended in ethyl acetate (5 mL), poured into a 5% solution of sodium bicarbonate (5 mL), brought to pH 7 with 1N HC1, and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate and filt~;red, and the solvent was removed under mild vacuum.
[004176] The resulting oils were treated to remove the BOC protective group, by dissolving the oil in 20 mL of methylene chloride (CH2C12), adding 4 mL of trifluoroacetic acid (TFA), and stirring the mixture for 10 min a~t 0°C. The CHZC12 and TFA were then removed under mild vacuum. The residue was concentrated from water twice, by heating under a vacuum, to remove any excess TFA, giving the unprotected derivative as a colorless syrup.
[000177) The resulting AAC-sulfonamide was puriiv3ed by preparative HPLC, using C~g reverse-phase ~,-Bondapack column, with an elution buffer containing 90% acetonitrile, 8% methanol, and 2% phosphate buffer, pH 7.~1, at 10 mL/min.
Impurities normally eluted within about 5 to 10 minutes, and product normally eluted after about 15 to 20 minutes.
[000178] Analysis to confirm the proper structure of A.AC-sulfonamide compound 1C was earned out, resulting in the following data: melting point:
140-1 °C;
IR (KBr), crri': 1170 (SOZsym), 1335 (SO2~), 1560 (amide II), 16.10 (amide I), (COOH); 3357 (NH, NH2); 1H-NMR (TFA), 300 MHz, 8, ppm: 3.30 (2H, s, CH2COOH);
3.42 (2H, s, CHZCONH); 7.25 (br s, 3H, CONH + S02NH2); ), 7.78 (d, 2H, AA'BB', 8.9), 7.91 (d, 2H, A.A'BB', 8.9), the NH and COOH protons are not seen in this solvent, being in fast exchange, 13C-NMR (TFA), 8, ppm: 26.8 (Me); 53.24 (CH2CH2); 53.70 (CHZCHZ); 60.18 (CHZCONH); 61.34 (CHZCOOH); 119.20 (C2iC3 -Ph), 126.5 (C3/C2 -Ph), 137.92 (C1/C4 -Ph), 142.39 (C4/C1 -Ph), 159.27 (CONH); 180.82 (COOH);
Anal., found: C, 41.62; H, 4.58; N, 14.47%; C~oH13N305S requires: C, 11.81; H, 4.56;
N, 14.63%.
EXAMPLE 5: PREPARATION OF BIS-SULFONAMIDES
[000179] Dianhydride versions of several of the AA.C reagents (including compound 4, EDTA; compound 5, DTPA; and compound 6, EG'TA) were used to prepare bis-sulfonamide compounds. This was done by adding 15 mmole; of the selected AAC
dianhydride reagent to a solution containing a double-molar excess (i.e., 30 mmole) of a selected sulfonamide reagent which had been dissolved in 100 mL of anhydrous dimethylformamide (DMF). The mixture was magnetically stirred at room temperature for 4 hours, then the reaction mixture was poured into 300 mL of methylene chloride (CH2Cl2). The solid condensate was filtered and thoroughly washed with CH2Cl2, then with acetone. HPLC purification was necessary in some cases, arid was done by elution with potassium phosphate buffer:methyl cyanide 2:1, v/v (10 mL/min), on a reversed-phase C 1 g Bondapack column.
[000180] Exemplary analytical data for two of the bis-sulfonamide compounds are provided below:
[000181] COMPOUND 4M2: m.p. 176-8 ~C (dec.); IR (KBr), cm': 1163 (S02Sym), 1356 (S02as), 1563 (amide II), 1600 (amide I), 1764 (COOH); 3330 (NH, NH2);
'H-NMR (D20-KOD), 300 MHz, S, ppm: 3.33 (4H, s, 2CH2COC>H); 3.54 (4H, t, CH2CH2); 3.60 (4H, s, 2CH2CONH); the S02NH2, CONH and COOH protons are not seen in this solvent mixture, being in fast exchange, '3C-NMR (D20-KOD), 8, ppm:
53.19 (CH2CH2); 53.76 (CH2C~i2); 60.23 (C~I2CONH); 61.13 (C'H2COOH); 159.3 (C-of thiadiazole); 170.5 (C-5 of thiadiazole); 176.28 (CONH); 180.70 (COOH);
elemental analysis: C14H2oNloOlosa.
[000182] COMPOUND SFz: m.p. 121-2 ~C (dec.); IR (KBr), cm'': 1173 (S02S'"r'), 1340 (S02as), 1560 (amide II), 1600 (amide I), 1760 (COOH); 3335 (NH, NH2);
1H-NMR (D20-KOD), 300 MHz, 8, ppm: 2.90 (t, 2H, CH2 of aminoethyl-benzenesulfonamide, 7.2), 3.47 (q, 2H, CH2 of aminoethylbenzenesulfonamide 6.5), 3.20 (4H, t, ethylenic CH2 near lateral nitrogens); 3.35 (4H, s, CH2 of the lateral acetates);
3.48 (4H, t, ethylenic CH2 near central nitrogen); 3.56 (4H, s, CFf2 of the acetamido groups); 3.91 (2H, s, CH2 of the central acetate); 7.42 (d, 2H, AA.~B ; 8.2), 7.75 (d, 2H, AA~B', 8.2), '3C-NMR (D20-KOD), 8, ppm: 36. 15 (C-10); 42.07 (C-9); 53.12 (C-4);
53.68 (C-3); 60.03 (C-2); 60.15 (C-7); 61.05 (C-5); 128.12 (C-13); 132.47 (C-12); 146.79 (C-14); 147.20 (C-11), 177.04 (C-8); 180.53 (C-1); 181.20 (C-6); elemental analysis:
C30H43N7012S2~
EXAMPLE 6: PREPARATION OF METAL COMPLEXES
[000183] A number of metal complexes (using zinc, copper, or aluminum) of some of the above-mentioned AAC-sulfonamide products were also prepared, and some were tested on rabbits. In general, these metal complexes were created by adding a strong alkaline agent (NaOH) to an aqueous solution of the AAC:-sulfonamide, to ensure that the very large majority of the carboxy groups would dissociate and form negatively-charged -COO- ionic groups. This was then added to a solution of a metallic salt (such as ZnCl2) which had been dissolved in water, and which contained ,gin abundance of free zinc (Zn~~, copper, or aluminim ions. The negatively-charged -COO- groups on the AAC-sulfonamide compounds formed ionic bonds with the positively-charged metal ions.
[000184] This general process is exemplified by compound SF2, as follows.
A suspension of 2 mmole (1.51 grams) of compound SFZ (full chemical name: 14-[4-(aminosulfonyl)phenyl]-3-[2-[[2-[4-(aminosulfonyl)-phenyl]-ethyl]amino]-2oxoethyl]-6,9-bis(carboxyrnethyl)-11-oxo-3,6,9,12-tetraazatetradecanoic arid) in 50 mL
water was treated with stoichiometric (2 mmole) 1N NaOH solution, in order to obtain the disodium carboxylate salt. The resulting salt solution was treated with a solution of ZnCl2 (1 mmole) in 5 mL of water, using NaOH to maintain the pH at 6.5 , The reaction was monitored by HPLC, on a stationary phase of Lichrospher 100 RP-18.5 Vim, with a 250x4 mm column packed by E. Merck, at 40°C. Isocratic elution with a premixed mobile phase (1 g octylamine was added to 100 mL of acetonitrile mixed with 900 mL of water) was performed. The eluent was buffered with phosphoric acid, maintaining the pH at 6, using a flow rate of 1.5 mL/min. After 4 h the solution was loaded onto an Amberlite XAD
1600 resin column (250 mL) and eluted with MeCN/water (1:10. v/v). The fractions containing the complex were evaporated to give a white solid of the metallic complex, with an overall yield of 95%.
[000185] Analytical data for the Zn-SFZ complex wc;re as follows: m.p.
>3000 C; IR(KBr), cm 1: 1170 (SOZsym}, 1336 (S028S), 1560 (am.ide II), 1610 (amide I), 1745 (COO'); 3335 (NHZ); elemental analysis: C3oH4,N~O,2SzZn, measured for Zn, C, H, and N.
EXAMPLE 7: ENZYME PREPARATIONS
[000186] Human CA-I and CA-II cDNAs were expressed in Escherichia coli strain BL21 (DE3), using the plasmids pACAIhCA-I and pA.CA/hCA-II; these two plasmids were a gift from Prof. Sven Lindskog, Umea Universitrr, Sweden, and are described in Lindskog et al 1991. Cell culture and expression conditions were as described in Behravan et al 1990. Enzymes were purified by affinity chromatography, using the method described Khalifah et al 1977. Enzyme concenmations were determined spectrophotometrically at 280 nm, utilizing a molar absorptivity of 49 mM''.crri' for CA-I and 54 mM'l.crri 1 for CA-II, respectively, based on Mr = 28.85 kDa for CA-I, and 29.30 kDa for CA-II, respectively (see Lindskog et al 1964 and Steiner et al 1975).
[000187] Bovine CA-IV was isolated from bovine hang microsomes, and its concentration was determined by titration with ethoxzolamide; see Maren et al 1993.
[000188] 4-nitrophenyl acetate (4-NPA) was used a;~ a color-generating substrate; when hydrolyzed by a CA enzyme, this substrate releases 4-nitrophenolate, an easily-visible compound which was measured for density at 400 nm, using a Cary spectrophotometer interfaced with an IBM compatible PC. Initial rates of 4-NPA
hydrolysis catalyzed by different CA isozymes were monitored. 'Typically, hydrolysis rates were measured for about 2 minutes, by which time they typically were approaching a saturation plateau.
[000189) Solutions of substrate were prepared in anhydrous acetonitrile; the substrate concentrations varied between 2.10'2 and 1.10'6 M, working at 25 0 C. A molar absorption coefficient of 18,400 M'~.crri 1 was used for the 4-nitr~~phenolate formed by hydrolysis, in the conditions of the experiments (pH 7.40), as reported in Pocker et al 1967. Non-enzymatic hydrolysis rates were always subtracted from observed rates.
Duplicate experiments were done for each inhibitor concentration; all values reported herein are the mean of such results.
[000190] Stock solutions ( 1 mM) of each candidate AAC-sulfonamide inhibitor compound were prepared in distilled and deionized water with 10 to 20% (v/v) DMSO; the DMSO was also tested by itself, and shown to be nom-inhibitory at the concentrations used. Dilutions up to 0.01 nM were done thereaft~;r with distilled-deionized water. Mixed solutions of enzymes and inhibitors were; preincubated together for 10 min at room temperature before the 4-NPA substrate was ;added, in order to allow the formation of enzyme-inhibitor complexes. The inhibition constant K, was determined as described by Pocker et al 1967. Enzyme concentrations were l2 nM for hCA-I, 3.5 nM
for hCA-II; higher concentrations (36 nM) were used to assay bC:A-IV activity, since the CA-IV isozyme from cows or humans is known to have lower esterase activity than CA-I
or CA-II isozymes.
[000191] The inhibition data which resulted from these tests, for compounds which were deemed to be potentially promising candidates, are provided in Table 1.
Complete inhibition data for all of the AAC-sulfonamide compounds that were created and tested, including those which did not perform effectively as ~~A
inhibitors and which are of no further interest with regard to this invention, are being :Filed as an appendix with this application. That appendix will be available for public inspe~~tion and copying upon issuance of a patent based on this application.
EXAMPLE 8: TESTS OF INTRA-OCULAR PRESSURES IN RABBITS
[000192] Adult male New Zealand albino rabbits weighing 3-3.5 kg were used in the experiments; three different animals were used for each inhibitor that was studied. All experimental procedures conformed to the Association for Research in Vision and Ophthalmology Resolution on the use of animals. The rabbits were kept in individual cages, with food and water provided ad libitum. The animals were maintained on a 12/12 hour light/dark cycle in a temperature controlled room, at 22-26~
C. Solutions or suspensions of inhibitors (2% by weight, as hydrochlorides or sodium carboxylate salts) were obtained in distilled deionized water. The pH of all AAC-sulfonamide compound solutions that were of interest were in the range of 6.~~ ~ 7.4; by contrast, the dorzolamide solution was at a more acidic pH, 5.8.
[000193] IOP was measured using a Digilab 30R pneumatonometer (BioRad, Cambridge, MA, USA), using procedures as described in articles such as Maren et al 1992 and Brechue et al 1993. The pressure readings were matched with two-point standard pressure measurements at least twice each day using a I)igilab Calibration verifier. All IOP measurements were done by the same investigator with the same tonometer. One drop of 0.2% oxybuprocaine hydrochloride (sold under the trademark NOVESINE, by Sandoz), diluted 1:1 with saline, was instilled ir.~ each eye within a few seconds before each set of pressure measurements. IOP was measured three times, at each time interval; the mean values are listed in the Tables herein. A
baseline IOP was immediately before administration of a candidate CA inhibitor dmg, then at 30 min after the instillation of the candidate drug, and again every 30 minutes, for a period of 4 to 6 hours. For all IOP experiments, one drop (50 ~.L) containing a candidate CA
inhibitor drug (at 2% concentration) was administered to one eye of a test animal, leaving the other eye as an untreated control. The ocular hypotensive activity values, as listed in the Tables, are expressed as the average difference in IOP values between the treated eye, and the control eye, for each animal; this was done to minimize a.ny diurnal, seasonal, or interindividual variations, as are commonly observed in rabbits (e.g., Maren et al 1992, Brechue et al 1993). All data are expressed as mean ~ standard error, using a one-tailed t test.
[000194] Unless otherwise indicated, all tests were clone on "normotensive"
rabbits, which had apparently healthy eyes and normal IOP values. Due to the absence of any disease or damage process in their eyes, these rabbits tend to provide more consistent and uniform data.
[000195] For tests involving "glaucomatous" rabbits, ocular hypertension was elicited in the right eye, by injecting an enzyme called alpha-chymotrypsin (purchased from Sigma-Aldrich), as described in Sugrue et al 1990.
Approximately four weeks were allowed to pass for the damage process to be fully manifested, and the IOP's of chymotrypsin-injected animals was checked. Animals having elevated IOP
values in the range of 30 to 36 mm Hg were designated as glaucomatous, and used in those tests.
[000196] The IOP-lowering potency data which resulted from these animal tests are provided in Table 3 (normotensive rabbits) and Table 4 (glaucomatous rabbits).
EXAMPLE 9: WATER SOLUBILITY AND SOLVENT PARTITIONING
[000197] The solubilities of various candidate AAC-sulfonamide drugs (in sodium salt form) in water was evaluated, using two different types of tests.
For comparison, dorzolamide was testing in the same manner, using :its commercially available HCl acid form, pH 5.8.
[000198] In the first type of test, a UV-calibrating standard solution of a candidate drug compound was prepared, by dissolving a precisely weighted amount (generally 1 mg) of the test compound, in a relatively large volume ( 10 mL) of methanol.
This mixture was then measured spectrophotometrically, using a Cary 3 spectrophotometer; if the initial measurement indicated a very strong absorption maximum, the solution was diluted to 1:10 by additional methanol. These initial tests determined: (i) the ultraviolet wavelength which was maximally absorbed by that particular compound, and (ii) the quantitative intensity of LJV light which was absorbed by that known concentration of the compound, at its maximal absorption wavelength, in excess methanol.
[000199] Once those data points were known for a certain test compound, a relatively large (excess) quantity of the test compound was placed in 10 mL of 0.039 M
phosphate buffered saline solution (pH 7.4). The saline solution was stirred for 3 hours at room temperature, to create a saline buffer solution which was saturated with the test compound. This saturated solution was then filtered through a Millipore 0.45 ~m filter, to remove any solid compound which had not dissolved in the saline buffer. The remaining saturated saline buffer was measured spectrophotometrically, using the predetermined UV wavelength for that compound. Total solubility of the satura~:ed saline buffer was then determined by the formula:
C' = C(A'/A) where C = concentration of the methanol standard solutia~n (mg/mL); A =
absorbance of the methanol standard solution; A' = absorbance of the saturated saline solution; C' = concentration of the saturated saline solution (mg/mL). The resulting aqueous saturation data for various compounds are provided in Table 2.
[000200] Solvent partition coefficients, using an aqueous solvent and an organic solvent, were also determined for various test compounds that were of interest, as follows. Equal volumes of chloroform and 0.1-ionic strength phosphate-buffered aqueous saline solution (pH 7.4) were mixed together, and a relatively small, nonsaturating quantity of a test compound was added to a mixture of saline andl chloroform.
The mixture was stirred at room temperature for 3 to 6 hours, until the test compound reached equilibrium between the organic and aqueous solvents. The concentration of the test compound in each phase was then measured, using UV spectrophotometry or HPLC
(e.g., Scozzafava et al, 1999a and 1999b).
EXAMPLE 10: IN VITRO CORNEAL PERMEATION TESTS
[000201] Since CA inhibitor drugs in topical eyedrops must be able to permeate into the eyeball to be effective, the ability of various AAC-sulfonamide test compounds to permeate into and through eye tissue was measured, using two different types of tests.
[000202] A first set of in vitro tests measured the ability of various test drugs to penetrate through the corneal tissue in the front of the eye, whcih is necessary for the drug to reach the clear liquids (humors) inside the eyeball. Transcorneal penetration was measured using the method of Maren et al 1983, with modifications as described in Pierce et al 1993 and Sharir et al 1994 for the HPLC assay of sulfonamides. In these tests, excised rabbit corneas, with either intact or denuded epithelium, were used, in saline buffer (pH 7.4). A piece of corneal tissue was placed across a 1.~ cm2 window between two chambers, each having a 6 mL volume. Concentrations of te;~t drugs ranging from 40 to 2000 pM were placed in the epithelial chamber (i.e., the chamber to which the epithelial side of the corneal tissue was exposed). At various intervals up to 4 hours after addition of a test drug to the chamber, samples of fluid were collected from the other (endothelial) chamber, which initially contained none of the test drug. The sampled fluids were assayed by HPLC methods (Pierce et al 1993; Sharir et al 1994), or enzymaticaliy (Maren et al 1983). The results of the drug analyses were used to calculate the rate constant of drug transfer across the cornea (k;"), by using the formula:
k;". (x 103 hr-') _ [drugc"aa]/[drug,~p;] x 60/t x 1000 where [druge"ao] = concentration of drug on endothelial side; [dmgep;] =
concentration of drug on epithelial side; and t = time, in minutes. The results of these in vitro tests for various compounds are included in Table 2. Except for test compound 1 O, the corneal permeation rates for the test compounds exceeded the permeation rates for dorzolamide.

EXAMPLE 11: DRUG DISTRIBUTION IN OCULAR TISSUES
[000203] In addition to the in vitro tissue permeation tests described in Example 10, an additional set of in vivo tests were also carried out, to measure the quantities of test compounds that were present in various different types of ocular tissues.
Because of their difficulty, and since each data point was gatherE;d by testing 3 different rabbits, these were carried out on only two test compounds, generally using procedures described in articles such as Maren et al 1992 and Brechue et al 1993.
[000204] In these tests, a known quantity a test drug; (a 2% solution in sodium salt form, in a single 50 ~,L eyedrop) was administered to both eyes of an animal.
Either 1 or 2 hours later, the animal was anesthetized and then sacrificed, using intracardiac air injection. The aqueous and vitreous humors were withdrawn using a hollow needle, and the cornea and anterior uvea (iris plus attached ciliary body) were removed and dissected, rinsed with distilled deionized water, blotted, weighed, and put into 1 to 2 mL of water. For isolation of the ciliary processes, intact anterior uvea rings were placed on a parafilm-covered piece of polystyrene foam in .a Petri dish.
The tissue was wetted with normal saline and dissected under a microscope. Ciliary processes were gently detached from the iris, cut, weighed, and put in 0.5 mL of distilled water. The tissue from 4 eyes (average weight of 8 mg/eye) was pooled for drug analysis.
[000205] Tissue samples containing the various different types of excised tissues were boiled for 5 minutes, in order to denature the carbonic anhydrase enzymes and release the test drug molecules from the enzyme-inhibitor complexes. Each resulting preparation was diluted, then incubated with a known quantity oi' fresh and functional CA
enzyme. The activity of the free enzyme in the presence of the inhibitor was determined as described above, and a calibration curve was used to determine the fractional inhibition of the enzyme by the test drug. This allowed drug concentrations to be measured in various different types of tissues.
[000206] As shown by the data in Table 6, relatively high concentrations of the candidate inhibitor compounds permeated into the ocular tissues and fluids, after topical administration of eyedrops containing these compounds.

Ocular tissue & fluid concentrations following corneal application of compounds 5M2 and 6M in normotensive rabbits Compound Time (h) Drug concentration (~,M) Cornea Aqueous humor Ciliary process SMZ 1 h 150 ~ 10 241 t 13 50 ~ 3 2h 45~5 39~3 19~1 6M 1 h 173 ~ 12 268 ~ 16 59 ~ 4 2h 66~S 53~3 37~3 EXAMPLE 12: SYNTHESIS OF ISOMERICALLY PURE END-CONNECTED
DTPA-MONO-SULFONAMIDE
[000207] This example describes a synthesis route that can be used, if desired, to create an AAC-mono-sulfonamide compound from DTPA, in substantially pure isomeric form, having a single sulfonamide-plus-ring substituent bonded to one of the "end-connected" carboxy groups of the DTPA residue. This reaction scheme is illustrated in FIG. 9. Similar methods also can be developed and used, if desired, for creating substantially pure isomeric forms of other mono-sulfomunide compounds from other candidate AAC reagents that have a combination of pendant carboxy groups with "substantively different" locations on an AAC reagent (such as a combination of "center-connected" and "end-connected" groups).
[000208] As shown in FIG. 9, the dianhydride version of DTPA (which has a single available carboxy group, in the center) can be treated, in a solvent such as diethyl ether, with a compound called diazomethane. This compound has a resonant structure, which can be drawn as either CHz=N=N or CHZ-N N; it will readily release its methyl group, which will combine with a free carboxy group to form a methyl ester group, which in this reaction will be located at the center-connected carboxy group, as shown in FIG. 9.

[000209] This first intermediate is then treated with water, to open the two anhydride rings and convert them into four "end-connected" carboxy groups.
[000210] This second intermediate is then mixed with a single molar equivalent of one of the sulfonamide reagents A through T. In Fi,g. 9, the variable "X"
represents either a nitrogen atom (for reagents A-P) or an oxygen atom (for reagents Q-T), and the variable "Y" represents the aromatic or unsaturated heterocyclic ring structure to which the sulfonamide group is bonded. This mixture is treated with a carbodiimide (such as di-isopropyl carbodiimide), in acetone solvent, to cause the same type of condensation reaction discussed in Examples 2 and 3 and shown in FIGS. 5 and 6. This condensation reaction will create either an amide bond (with sulfonamide compounds A-P) or an ester bond (with sulfonamide compounds Q-T) between the methylated AAC
intermediate and the sulfonamide reagent. Since the four unsubstituted carboxy groups on the center-methylated DTPA derivative are all equivalent, the pr<;dominant product from this condensation reaction with a single molar equivalent of the sulfonamide reagent will be a mono-sulfonamide compound. If any di-, tri-, or tetra-sulfonamide compounds are created, they can be easily removed from the mono-sulfonamide product, by using preparative chromatography or other known purification methods, since their molecular weights will be substantially different from the desired mono-sulfonamide product.
[000211] If removal of the methyl group from the center-connected methyl-ester group is desired (to recreate a free carboxy group), the condensation product can be treated with acid in the presence of a suitable ion exchange resin (such as one of the "condensate polishing" versions of DOWER, sold by Dow Chemical Company). This presumably should be done prior to chromatography or other purification steps.
[000212] These will generate a substantially pure single-isomer preparation of a DTPA-mono-sulfonamide compound, having the sulfonamide group and its ring structure bonded to an end-connected carboxy residue, as shown in FIG. 9.
EXAMPLE 13: SYNTHESIS OF ISOMERICALLY PURE CENTER-CONNECTED
DTPA-MONO-SULFONAMIDE
OOf 02131 This example describes a synthesis route that can be used, if desired, to create an AAC-mono-sulfonamide compound from D'TPA, in substantially pure isomeric form, having a single sulfonamide-plus-ring substituent bonded to the "center-connected" carboxy group of the DTPA residue. This reaction scheme is illustrated in FIG. 10. Similar methods also can be developed and used, if desired, for creating substantially pure isomeric forms of other mono-sulfonamide compounds from other candidate AAC reagents that have a combination of pendant carboxy groups in different locations on the AAC reagent.
OOf 02141 In this method, a halogenated tertiary amine, having two methyl-ester groups coupled (through C2 or other lower alkyl chains) to the tertiary amine, is used. This halogenated di(methylester) amine, if not commercially available, can be created by steps such as shown at the top of FIG. 10, using ethylene oxide (also known as dioxirane) to convert a secondary di(methylester) amine into a tertiary amine with an alcohol group. The hydroxy group on the alcohol-amine-diester is then displaced with a chlorine atom, by using an agent such as thionyl chloride (SOCIz).
OOf 02151 A double-molar ratio of the resulting intermediate is then reacted with amino-acetic acid (which is glycine, the common amino acid) under conditions which displace both of the hydrogen atoms on the amino group o~f the glycine, and the chlorine atoms on two molecules of the chloroalkyl-amine-diester compound.
This condensation reaction creates a tetra-methyl-ester protected intermediate, having a single free carboxy group (contributed by the glycine) located in the center of the tetra-methylated DTPA derivative, as shown in FIG. 10.
[000216] This intermediate is then reacted with a single molar equivalent of a sulfonamide-plus-ring reagent, such as one of the reagents A-T shown in FIGS. 2A and 2B, in acetone solvent, with a carbodiimide present. This leads to a condensation reaction in the same manner described in Examples 2 and 3 and shown in FIGS. 4 and 6, in which the sulfonamide reagent bonds to the sole unprotected carboxy group in the center of the tetra-methylated DTPA derivative, forming an amide bond (reagents A-P) or an ester bond (reagents Q-T). The four methyl protective groups can then be removed from the sulfonamide condensate, by using a condensate-polishing DOWI:X resin under acid conditions.
[000217] As shown in FIG. 10, this will create a mono-sulfonamide DTPA
derivative, with the sulfonamide bonded to the central carboxy. In the final product shown in FIG. 10, the variable "X" represents either a nitrogen atom (for reagents A-P) or an oxygen atom (for reagents Q-T), and the variable "Y" represents the aromatic or unsaturated heterocyclic ring structure to which the sulfonamide group is bonded.
Preparative HPLC or similar purification steps can be used to remove any reaction byproducts from the desired isomerically pure DTPA-mono-sulfbnamide product.
[000218] Thus, there has been shown and described a new and useful class of water-soluble carbonic anhydrase inhibitor drugs that are higr~ly effective in reducing intraocular pressure and treating glaucoma, when administered in non-acidic eyedrop formulations. Although this invention has been exemplified for purposes of illustration and description by reference to certain specific embodiments, it will be apparent to those skilled in the art that various modifications, alterations, and equivalents of the illustrated examples are possible. Any such changes which derive directly from the teachings herein, and which do not depart from the spirit and scope of the invention, are deemed to be covered by this invention.
[000219] In view of the above, it will be seen that the several advantages of the invention are achieved and other advantageous results attained.
[000220] As various changes could be made in the above methods and compositions without departing from the scope of the invention, it is intended that all matter contained in the above description be interpreted as illusb~ative and not in a limiting sense.
[000221] All references cited in this specification are hereby incorporated by reference. The discussion of the references herein is intended mexely to summarize the assertions made by their authors and no admission is made that any reference constitutes prior art. Applicant reserves the right to challenge the accuracy and pertinency of the cited references.
REFERENCES
Behravan, G., et al, "Fine tuning of the catalytic properties of carbonic anhydrase:
Studies of a Thr200-His variant of human isoenzyme II," Eur. J. Biochem. 190:

( 1990) Brechue, W.F., et al, "pH and drug ionization affects ocular pressure lowering of topical carbonic anhydrase inhibitors. Invest. Ophthalmol. Vis. Sci., 1993, 34, 2581-2587.
Casini, A., et al, "Carbonic anhydrase inhibitors: Water soluble 4-sulfamoylphenyl-thioureas as topical intraocular pressure lowering agents with long lasting effects," J. Med. Chem. 43: 4884-4892 (2000) Chen, H.H., et al, "2H-Thieno [3,2-a]- and [2,3-a]-1,2-thiazine-6-sulfonamide 1,1-dioxides as ocular hypotensive agents: synthesis, carbonic anhydrase inhibition and evaluation in the rabbit," Bioorg. Med. Chem. 8: 957-975 (2000) Cingolani, E., "Sulla alogenazione della p-amino-benzene-solfonamide (derivatialogenati nucleari)," Gazz. Chim. Ital. 78: 275-282 (1948) Crippa, G.B., et al., "Derivati solfonamidici del pirazolo," Gazz. Chim. Ital.
71: 97-99 (1941) Eller, M.G., et al., "Topical carbonic anhydrase inhibitors: III.
Optimization model for corneal penetration of ethoxzolamide analogues,"
J. Pharm. Sci. 74: 155-160 (1985) Jitianu, A., et al., "Complexes with biologically active ligands. Part 8.
Synthesis and carbonic anhydrase inhibitory activity of 5-benzoylamido- and 5-(3-nitrobenzoylamido)-1,3,4-thiadiazole-2-sulfonamide and their metal complexes,"
Main Group Met. Chem. 20: 151-156 (1997).
Khalifah, R.G., et al, "Carbon-13 nuclear magnetic resonance probe of active site ionization of human carbonic anhydrase B," Biochemistry 16: 22.41-2247 (1977) Lindskog, S., et al, "The catalytic mechanism of carbonic anhydrase" Proc.
Natl.
Acad Sci. USA 70: 2505-2508 (1964) Lindskog, S., et al, "Structure-function relations in human carbonic anhydrase II
as studied by site-directed mutagenesis," pp. 1-13 in Carbonic A~:hydrase 0 From Biochemistry and Genetics to Physiology and Clinical Medicine (Botre, F., et al, eds.
(VCH, Weinheim, (1991) Maren, T.H., et al, "The transcorneal permeability of sulfonamide carbonic anhydrase inhibitors and their effect on aqueous humor secretion," Exp. Eye Res. 36:
457-480 (1983) Maren, T.H., et al, "Relations among IOP reduction, ocular disposition and pharmacology of the carbonic anhydrase inhibitor ethoxzolamide," Exp. Eye Res.
55: 73-79 ( 1992) Pierce, W.M., et al, "Topically active ocular carbonic anlrydrase inhibitors:
Novel biscarbonylamidothiadiazole sulfonamides as ocular hypotensivE; agents," Proc.
Soc. Exp.
Biol. Med. 203: 360-365 (1993) Pocker, Y., et al, "The catalytic versatility of erythrocyte carbonic anhydrase: III.
Kinetic studies of the enzyme-catalyzed hydrolysis of p-nitrophenyl acetate,"
Biochemistry 6: 668-678 ( 1967) Scozzafava, A et al., "Carbonic anhydrase inhibitors: Novel compounds containing S-NH moieties: Sulfenamido-sulfonamides, sulfenim:ido-sulfonamides and their interaction with isozymes I, II and IV," J. Enzyme Inhib. 13 : 419-442 ( 1998).
Scozzafava, A., et al, "Carbonic anhydrase inhibitors: Synthesis of water-soluble, topically effective intraocular pressure lowering aromatic/heterocyclic sulfonamides containing cationic or anionic moieties: is the tail more important than the ring?" J. Med.
Chem. 42: 2641-2650 (1999) Scozzafava, A., et al, "Carbonic anhydrase inhibitors: Synthesis of water-soluble, amino acyl/dipeptidyl sulfonamides possessing long-lasting intra.ocular pressure-lowering properties via the topical route. J. Med. Chem. 42: 3690-3700 (1!99) Scozzafava, A., et al, "Carbonic anhydrase inhibitors. Perfluoroalkyl/aryl-substituted derivatives of aromatic/ heterocyclic sulfonamides as topical intraocular pressure lowering agents with prolonged duration of action," J. cited. Chem.
43: 4542-4551 (2000) Sharir, M., et al, "Pharmacokinetics, acid-base balance and intraocular pressure effects of ethyloxaloylazolamide, a novel topically active carbonic anhydrase inhibitor,"
Exp. Eye Res. 58: 107-116 (1994) Steiner, H., et al, "The catalytic mechanism of carbonic anhydrase: Hydrogen-isotope effects on the kinetic parameters of the human C isoenzyme," Eur. J.
Biochem.
59: 253-259 (1975) Sugrue, M.F., et al, "L-662,583 is a topically effective ocular hypotensive carbonic anhydrase inhibitor in experimental animals," Br. J. Ph~xrmacol. 99:

( 1990) Supuran, C.T. et al., "Novel aromatic/heterocyclic sulfonamides and their metal 7_ complexes as inhibitors of carbonic anhydrase isozymes I, II and. IV" J.
Enryme Inhib.
12:37-51 (1997) _78_

Claims (2)

1. A carbonic anhydrase inhibitory compound or a salt thereof, wherein the compound has a formula ZLn;

n is 1 or 2;
Z is selected from the group consisting of:

R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and N(R16)-;
X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C 1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the grout' consisting of C1-4 alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the ,group consisting of C1-3 alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
L is selected from the group consisting of:

Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:

the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;

R22, R26 and R28 are independently selected from the group consisting of -NH-, -O-, and -S-;

R23 is selected from the group consisting of -N=, -NHCH=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;

R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazine, and C 1-3-alkyl, wherein:

the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;

R25 is selected from the group consisting of C1-3-alkyl and -(CH2)pC(O)NH-, wherein p is from zero to 2.

2. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 1, wherein 2 is selected from the group consisting of:

and L is selected from the group consisting of:

X is F, Cl, Br, or I;

6. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 1, which has at least one of (a) a greater aqueous solubility at pH 7 than that of dorzolamide or brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of dorzolamide or brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of an amount of brinzolamide providing equal peak efficacy when administered to an eye of a mammal.

7. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 1, which has an aqueous solubility at pH 7 of at least about 1% w/w.

8. A carbonic anhydrase inhibitory compound or salt thereof according to claim 1, which inhibits a mammalian carbonic anhydrase isozyme II at a K I of about 15 nM or less.

9. A carbonic anhydrase inhibitory compound or salt thereof according to claim 1, which inhibits a mammalian carbonic anhydrase isozyme IV at a K I of about 50 nM or less.

10. A carbonic anhydrase inhibitory compound or salt thereof according to claim 1, which produces a decrease in intraocular pressure of at least 3 mm Hg for a duration of at least 4 hours in normotensive rabbits upon ocular application at a concentration of 2% (w/w) in aqueous eyedrops.

11. A carbonic anhydrase inhibitory compound or a salt thereof, wherein the compound has a formula ZL n;

n is 1 or 2;

Z is selected from the group consisting of:

R1 is hydroxy when n is 1;

R1 is a bond when n is 2;

R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;

R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;

X1 is selected from the group consisting of a bond, -O-, -S-, and -N(R16)-;

X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;

X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;

X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;

X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;

X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;

R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;

R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;

R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;

R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;

L is selected from the group consisting of:

Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:

the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;

Y3 is selected from the group consisting of hydrogen and sulfonamide;

Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:

the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;

R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;

R22, R26 and R28 are independently selected from the group consisting of -NH-, -O-, and -S-;

R23 is selected from the group consisting of N=, -NHCH=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;

R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:

the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;

R25 is selected from the group consisting of C1-3-alkyl and -(CH2)pC(O)NH-, wherein p is from zero to 2; and wherein if Z is X1 is N-, R2 and R3 are each -CH2-, R5 and R6 are -carboxymethyl, and R7 and R8 is C2 alkyl, ZL n consists of ZL and the compound is in a substantially isomerically purified form of greater than 80% (w/w) one isomer.

12. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 11, wherein Z is selected from the group consisting of:

and L is selected from the group consisting of:

13. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 11 wherein the salt comprises at least one ion selected from the group Zn+2, Al+2, and Cu+2 ion.

14. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 11 which has a log chloroform/water partition coefficient (Log P) of from about 0.2 to about 2Ø

15. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 14 which has a Log P of from about 1.0 to about 2Ø

16. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 11, which has at least one of (a) a greater aqueous solubility at pH 7 than that of dorzolamide or brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of dorzolamide or brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of an amount of brinzolamide providing equal peak efficacy when administered to an eye of a mammal.

17. A carbonic anhydrase inhibitory compound or a salt thereof according to claim 11, which has an aqueous solubility at pH 7 of at least about 1% w/w.

18. A carbonic anhydrase inhibitory compound or salt thereof according to claim 11, which inhibits a mammalian carbonic anhydrase isozyme II at a K I of about 15 nM or less.

19. A carbonic anhydrase inhibitory compound or salt thereof according to claim 11, which inhibits a mammalian carbonic anhydrase isozyme IV at a K I of about 50 nM or less.

20. A carbonic anhydrase inhibitory compound or salt thereof according to claim 11, which produces a decrease in intraocular pressure of at least 3 mm Hg for a duration of at least 4 hours in normotensive rabbits upon ocular application at a concentration of 2% (w/v) in aqueous eyedrops.

21. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor, the method comprising reacting in the presence of water and a coupling agent, compound A with compound B to produce a reaction product, wherein compound A is R is C1-7 alkyl, and wherein compound B is selected from the group consisting of:

X is F,Cl, Br, or I;

and hydrolyzing the reaction product, thereby removing the C1-5 alkyl and producing the isomerically pure form of the carbonic anhydrase inhibitor.

22. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor according to claim 21, wherein the coupling agent is a carbodiimide coupling agent.

23. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor according to claim 21, wherein the carbodiimide coupling agent is selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and diisopropyl carbodiimide.

24. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor, the method comprising reacting in the presence a coupling agent, compound A with compound B to produce a reaction product, wherein compound A is R1, R2, R3, and R4 are each independently C1-7 alkyl, and wherein compound B is selected from the group consisting of and hydrolyzing the reaction product, thereby removing the C1-7 alkyl and producing the isomerically pure form of the carbonic anhydrase inhibitor.

25. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor according to claim 34, wherein the coupling agent is a carbodiimide coupling agent.

26. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor according to claim 21, wherein the carbodiimide coupling agent is selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and diisopropyl carbodiimide.

27. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor, the method comprising reacting in the presence a coupling agent, compound A with compound B to produce a reaction product, wherein compound A is R is C1-7 alkyl, and wherein compound B is selected from the group consisting of and hydrolyzing the reaction product, thereby removing the C1-7 alkyl and producing the isomerically pure form of the carbonic anhydrase inhibitor.
28. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor according to claim 27, wherein the coupling agent is a carbodiimide coupling agent.
29. A method of preparing a substantially isomerically pure form of a carbonic anhydrase inhibitor according to claim 28, wherein the carbodiimide coupling agent is selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and diisopropyl carbodiimide.
30. A method of preparing a carbonic anhydrase inhibitory compound, the method comprising reacting in the presence of a coupling agent, a compound A
selected from the group consisting of:
R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and -N(R16)-;
X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
with a sulfonamide compound B selected from the group consisting of Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazine, and C1-3-alkyl, wherein:

the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;
R22, R26 and R28 are independently selected from the group consisting of NH-, -O-, and -S-;
R23 is selected from the group consisting of-N=, -NHCH(=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;
R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C1-3-alkyl and -(CH2)pC(O)NH-, wherein p is from zero to 2.
31. A method of preparing a carbonic anhydrase inhibitory compound according to claim 30, wherein the coupling agent is a carbodiimide coupling agent.
32. A method of preparing a carbonic anhydrase inhibitory compound according to claim 31, wherein the carbodiimide coupling agent is selected from the group consisting of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 1,3-diisopropyl carbodiimide.
33. A method of preparing a carbonic anhydrase inhibitory compound according to claim 30 wherein compound A is selected from the group consisting of:
and wherein compound B is selected from the group consisting of:
X is F, Cl, Br, or I;

34. A method of preparing a carbonic anhydrase inhibitory compound, the method comprising reacting in the presence of a coupling agent a compound A
and a compound B, wherein compound A is selected from the group consisting of:
R1 is hydroxy when n is 1;
R1 is a bond when n is 2;

R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and -N(R16)-;
X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and- N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C 1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
with a sulfonamide compound B selected from the group consisting of:
Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;
R22, R26 and R28 are independently selected from the group consisting of -NH-, -O-, and -S-;
R23 is selected from the group consisting of -N=, -NHCH=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;
R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C1-3-alkyl and -(CH2)p C(O)NH-, wherein p is from zero to 2; and wherein if compound A is X1 is N-, R2 and R3 are each -CH2-, R5 and R6 are -carboxymethyl, and R7 and R8 is C2 alkyl.
35. A method of preparing a carbonic anhydrase inhibitory compound according to claim 34 wherein compound A is selected from the group consisting of:
and wherein compound B is selected from the group consisting of:
36. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound having a formula ZL n wherein n is 1 or 2, Z is selected from the group consisting of:

R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and N(R16)-;
X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the ,group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
L is selected from the group consisting of:
Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazine, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazine, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;
R22, R26 and R28 are independently selected from the group consisting of -NH-, -O-, and -S-;
R23 is selected from the group consisting of N=, -NHCH=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;
R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazine, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C1-3-alkyl and -(CH2)pC(O)NH-, wherein p is from zero to 2.

37. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 36, wherein Z is selected from the group R = bond or hydroxy; and L is selected from the group consisting of:
X is F, Cl, Br, or I;
38. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37 wherein the salt comprises at least one ion selected from the group Zn+2, Al+2, and Cu+2 ion.

39. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37 wherein the carbonic anhydrase inhibitory compound has a log chloroform/water partition coefficient (Log P) of from about 0.2 to about 2Ø

40. The method of treating or preventing ocular hypertension and/or glaucoma according to claim 39 wherein the carbonic anhydrase inhibitory compound has a Log P
of from about 1.0 to about 2Ø

41. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37, wherein the compound has at least one of (a) a greater aqueous solubility at pH 7 than that of dorzolamide or brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of dorzolamide or brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of an amount of brinzolamide providing equal peak efficacy when administered to an eye of a mammal.

42. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37, wherein the compound has an aqueous solubility at pH 7 of at least about 1% w/w.

43. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37, wherein the compound inhibits a mammalian carbonic anhydrase isozyme II at a K I of about 15 nM or less.

44. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37, wherein the compound inhibits a mammalian carbonic anhydrase isozyme IV at a K I of about 50 nM or less.

45. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 37, wherein the compound or salt produces a decrease in intraocular pressure of at least 3 mm Hg for a duration of at least 4 hours in :normotensive rabbits upon ocular application at a concentration of 2% (w/w) in aqueous eyedrops.

46. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound having a formula ZL n wherein n is 1 or 2; wherein Z is selected from the group consisting of:
R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and -N(R16)-;
X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;

R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
L is selected from the group consisting of:
Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;
R22, R26 and R28 are independently selected from the group consisting of NH-, -O-, and -S-;
R23 is selected from the group consisting of -N=, -NHCH=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;
R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C1-3-alkyl and -(CH2)pC(O)NH-, wherein p is from zero to 2;
and wherein if Z is ZL n consists of ZL and the compound is in a substantially isomerically purified form of greater than 80% (w/w) one isomer.

47. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46 wherein the substantially isomerically purified form is at least 90%
(w/w) one isomer.

48. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46, wherein Z is selected from the group consisting of:
L is selected from the group consisting of:

X is F, Cl, Br, or I;

49. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46 wherein the salt comprises at least one ion selected from the group Zn+2, Al+2, and Cu+2.

50. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46 wherein the carbonic anhydrase inhibitory compound has a log chloroform/water partition coefficient (Log P) of from about 0.2 to about 2Ø

51. The method of treating or preventing ocular hypertension and/or glaucoma according to claim 46 wherein the carbonic anhydrase inhibitory compound has a Log P
of from about 1.0 to about 2Ø

52. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46, wherein the compound has at least one of (a) a greater aqueous solubility at pH 7 than that of brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of 1%
(w/w) brinzolamide aqueous eyedrops administered to an eye of a mammal.

53. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46, wherein the compound has an aqueous solubility at pH 7 of at least about 1% w/v.

54. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46, wherein the compound inhibits a mammalian carbonic anhydrase isozyme II at a K1 of about 15 nM or less.

55. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46, wherein the compound inhibits a mammalian carbonic anhydrase isozyme IV at a K1 of about 50 nM or less.

56. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 46, wherein the compound or salt produces a decrease in intraocular pressure of at least 3 mm Hg for a duration of at least 4 hours in normotensive rabbits upon ocular application at a concentration of 2% (w/w) in aqueous eyedrops.

57. An ophthalmic composition comprising a carbonic anhydrase inhibitory compound or a pharmaceutically acceptable salt thereof, in an opthalmically acceptable vehicle, said compound having a formula ZL n wherein n is 1 or 2, Z is selected from the group consisting of:

R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C 1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and -N(R16)-;
X2 is selected from the group consisting of -O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
L is selected from the group consisting of:

Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, brume, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;
R22, R26 and R28 are independently selected from the group consisting of -NH-, -O-, and -S-;
R23 is selected from the group consisting of -N=, -NHCH=, -NHCH2CH=, -NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;
R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C1-3-alkyl and -(CH2)pC(O)NH-, wherein p is from zero to 2.

58. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57, wherein Z is selected from the group consisting of:

L is selected from the group consisting of:

X is F, Cl, Br, or I;

59. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57 wherein the salt comprises at least one ion selected from the group Zn+2, Al+2, and Cu+2.

60. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57 wherein the carbonic anhydrase inhibitory compound has a log chloroform/water partition coefficient (Log P) of from about 0.2 to about 2Ø

61. The method of treating or preventing ocular hypertension and/or glaucoma according to claim 60 wherein the carbonic anhydrase inhibitory compound has a Log P

of from about 1.0 to about 2Ø

62. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57, wherein the compound has at least one of (a) a greater aqueous solubility at pH 7 than that of brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of 1 %
(w/w) brinzolamide aqueous eyedrops administered to an eye of a mammal.

63. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57, wherein the compound has an aqueous solubility at pH 7 of at least about 1 % w/v.

64. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57, wherein the compound inhibits a mammalian carbonic anhydrase isozyme II at a K1 of about 15 nM or less.

65. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57, wherein the compound inhibits a mammalian carbonic anhydrase isozyme IV at a K1 of about 50 nM or less.

66. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 57, wherein the compound or salt produces a decrease in intraocular pressure of at least 3 mm Hg for a duration of at least 4 hours in normotensive rabbits upon ocular application at a concentration of 2% (w/w) in aqueous eyedrops.

67. An ophthalmic composition comprising a carbonic anhydrase inhibitory compound or a pharmaceutically acceptable salt thereof, in an ophthalmically acceptable vehicle, said compound having a formula ZL n wherein n is 1 or 2; wherein Z is selected from the group consisting of:

R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and N(R16)-;
X2 is selected from the group consisting of-O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
L is selected from the group consisting of:

Y1 and Y2 are independently selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
Y3 is selected from the group consisting of hydrogen and sulfonamide;
Y4 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino and thiol;
R21 and R27 are independently selected from the group consisting of a bond and C1-2-alkyl;
R22, R26 and R28 are independently selected from the group consisting of -NH-, -O-, and -S-;

R23 is selected from the group consisting of -N=, -NHCH=, -NHCH2CH=, NH(CH2)2CH=, -OCH=, -OCH2CH=, -O(CH2)2CH=, -SCH=, -SCH2CH=, and -S(CH2)2CH=;
R24 is selected from the group consisting of hydrogen, chloro, bromo, fluoro, iodo, cyano, nitro, amino, sulfonamide, hydroxy, thiol, carboxy, hydrazino, and C1-3-alkyl, wherein:
the C1-3-alkyl is optionally substituted with a substituent selected from the group consisting of hydroxy, amino, and thiol;
R25 is selected from the group consisting of C1-3-alkyl and -(CH2)p C(O)NH-, wherein p is from zero to 2.
and wherein if L is ZL n consists of ZL and the compound is in substantially isomerically purified form of greater than 80% (w/w) one isomer.

68. An ophthalmic composition according to claim 90, which has a pH of from about 6.6 to about 7.8.

69. An ophthalmic composition according to claim 90 wherein Z of the carbonic anhydrase inhibitory compound is selected from the group:

and L is selected from the group consisting of:

X is F, Cl, Br, or I;

70. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67 wherein the salt comprises at least one ion selected from the group Zn+2, Al+2, and Cu+2.

71. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67 wherein the carbonic anhydrase inhibitory compound has a log chloroform/water partition coefficient (Log P) of from about 0.2 to about 2Ø

72. The method of treating or preventing ocular hypertension and/or glaucoma according to claim 67 wherein the carbonic anhydrase inhibitory compound has a Log P
of from about 1.0 to about 2Ø

73. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67, wherein the compound has at least one of (a) a greater aqueous solubility at pH 7 than that of brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of 1 %
(w/w) brinzolamide aqueous eyedrops administered to an eye of a mammal.

74. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67, wherein the compound has an aqueous solubility at pH 7 of at least about 1 % w/v.

75. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67, wherein the compound inhibits a mammalian carbonic anhydrase isozyme IV at a K I of about 15 nM or less.

76. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67, wherein the compound inhibits a mammalian carbonic anhydrase isozyme IV at a K I of about 50 nM or less.

77. A method of treating or preventing ocular hypertension and/or glaucoma according to claim 67, wherein the compound or salt produces a decrease in intraocular pressure of at least 3 mm Hg for a duration of at least 4 hours in normotensive rabbits upon ocular application at a concentration of 2% (w/w) in aqueous eyedrops.

78. A carbonic anhydrase inhibitory compound or a salt thereof, the compound having a formula ZL n wherein n is 1 or 2, Z is selected from the group consisting of:

R1 is hydroxy when n is 1;
R1 is a bond when n is 2;
R2 and R3 are independently selected from the group consisting of a bond, -CH2-, and -(CH2)2-;
R5 and R6 are each independently selected from the group consisting of hydrogen, carboxy, hydroxyphenyl, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
X1 is selected from the group consisting of a bond, -O-, -S-, and N(R16)-;
X2 is selected from the group consisting of-O-, -S-, and -N(R17)-;
X3 is selected from the group consisting of -O-, -S-, and -N(R18)-;
X4 is selected from the group consisting of -O-, -S-, and -N(R19)-;
X5 is selected from the group consisting of -O-, -S-, and -N(R20)-;
X6 is selected from the group consisting of -O-, -S-, and -N(R21)-;
R4, R16, R17, R18, R19, R20 and R21 are independently selected from the group consisting of hydrogen, carboxy, and C1-2-alkyl, wherein the C1-2-alkyl is optionally substituted with carboxy;
R7 and R8 are independently selected from the group consisting of C1-5-alkyl, wherein the sum of the carbon atoms in R7 and R8 is from 2 to 6;
R9, R10 and R11 are independently selected from the group consisting of C1-4-alkyl, wherein the sum of the carbon atoms in R9, R10 and R11 is from 3 to 6;
R12, R13, R14 and R15 are independently selected from the group consisting of C1-3-alkyl, wherein the sum of the carbon atoms in R12, R13, R14 and R15 is from 4 to 6;
and L is selected from the group consisting of -NH-M-SO2NH2, -N = M-SO2NH2, -O-M-SO2NH2, and -NH-NH-M-SO2NH2 wherein M is selected from the group consisting of pyranyl, pyridinyl, pyrrolyl, pyrimidinyl, thiazolyl, thiophenyl, thiolanyl, imidazolyl, oxazolyl, isoxazolyl, and furanyl.

79. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein Z is selected from the group consisting of:

80. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein the salt comprises at least one ion selected from the group Zn+2, Al+2, and Cu+2.

81. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein the carbonic anhydrase inhibitory compound has a log chloroform/water partition coefficient (Log P) of from about 0.2 to about 2Ø

82. A carbonic anhydrase inhibitory compound or a salt according to claim 81, wherein the carbonic anhydrase inhibitory compound has a Log P of from about 1.0 to about 2Ø

83. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein the compound has at least one of (a) a greater aqueous solubility at pH 7 than that of brinzolamide, (b) a greater inhibitory activity against at least one of mammalian carbonic anhydrase isozymes II or IV than that of brinzolamide, or (c) a longer duration of intraocular pressure-reducing activity in a mammal, when formulated in aqueous eyedrops and administered to an eye of a mammal, than that of 1 % (w/w) brinzolamide aqueous eyedrops administered to an eye of a mammal.

84. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein the compound has an aqueous solubility at pH 7 of at least about 1 %
w/w.

85. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein the compound inhibits a mammalian carbonic anhydrase isozyme II at a K1 of about 15 nM or less.

86. A carbonic anhydrase inhibitory compound or a salt according to claim 78, wherein the compound inhibits a mammalian carbonic anhydrase isozyme IV at a K1 of about 50 nM or less.

87. An amine-carboxy-sulfonamide compound or salt thereof, comprising:
a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component hawing at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the compound is administered topically in an aqueous eyedrop formulation to a mammalian eye.

88. The amine-carboxy-monosulfonamide compound or salt of claim 87 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

89. The amine-carboxy-monosulfonamide compound or salt of claim 87 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normatensive rabbits by at least 3 mm Hg for at least four hours after administration.

90. An amine-carboxy-sulfonamide compound or salt thereof, comprising:
a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;
b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

91. The amine-carboxy-monosulfonamide compound or salt of claim 90 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

92. The amine-carboxy-monosulfonamide compound or salt of claim 90 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

93. An amine-carboxy-sulfonamide compound or salt thereof, comprising:
a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;
b. an amine-carboxy-alkyl component, having at least one; C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

94. The amine-carboxy-monosulfonamide compound or salt of claim 93 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

95. The amine-carboxy-monosulfonamide compound or salt of claim 93 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

96. The amine-carboxy-monosulfonamide compound or salt of claim 93 wherein the substantially isomerically pure compound or salt is at least about 90% one isomer of total weight.

97. An amine-carboxy-disulfonamide compound or salt thereof, comprising:
a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component hawing at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered to a mammalian eye.

98. The amine-carboxy-monosulfonamide compound or salt of claim 97 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

99. The amine-carboxy-monosulfonamide compound or salt of claim 97 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

100. An amine-carboxy-sulfonamide compound or salt thereof, comprising:
a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;
b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-disulfonamide compound hay, sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

101. The amine-carboxy-monosulfonamide compound or salt of claim 100 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

102. The amine-carboxy-monosulfonamide compound or salt of claim 100 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

103. An amine-carboxy-sulfonamide compound or salt thereof, comprising:
a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;
b. an amine-carboxy-alkyl component, having at least one; C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

104. The amine-carboxy-monosulfonamide compound or salt of claim 103 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

105. The amine-carboxy-monosulfonamide compound or salt of claim 103 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

106. An aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:
a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component hawing at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

107. The aqueous eyedrop formulation of claim 106 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

108. The aqueous eyedrop formulation of claim 107 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

109. An aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:
a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;
b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which. can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

110. The aqueous eyedrop formulation of claim 109 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

111. The aqueous eyedrop formulation of claim 109 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

112. An aqueous eyedrop formulation comprising an a mine-carboxy-sulfonamide compound or salt therof, the compound having:
a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;
b. an amine-carboxy-alkyl component, having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

113. The aqueous eyedrop formulation of claim 112 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

114. The aqueous eyedrop formulation of claim 112 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

115. An aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:
a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

116. The aqueous eyedrop formulation of claim 115 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

117. The aqueous eyedrop formulation of claim 115 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

118. An aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:
a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

119. The aqueous eyedrop formulation of claim 118 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

120. The aqueous eyedrop formulation of claim 118 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

121. An aqueous eyedrop formulation comprising an canine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component, having at least one; C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

122. The aqueous eyedrop formulation of claim 121 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

123. The aqueous eyedrop formulation of claim 121 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

124. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound comprising an amine-carboxy-monosulfonamide having:

a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the compound is administered topically in an aqueous eyedrop formulation to a mammalian eye.

125. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 124 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

126. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 124 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

127. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound comprising an amine-carboxy-monosulfonamide having:

a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

128. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 127 wherein the compound or salt, when formulated in a 2% (w/v) aque28ous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

129. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 127 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

130. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound comprising an amine-carboxy-monosulfonamide having:

a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component, having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

131. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 130 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

132. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 130 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

133. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering; ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound comprising an amine-carboxy-monosulfonamide having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component hawing at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered to a mammalian eye.

134. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 133 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

135. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 133 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

136. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound comprising an amine-carboxy-monosulfonamide having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

137. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 136 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

138. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 136 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

139. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, a carbonic anhydrase inhibitory compound or an a salt thereof in an amount effective in treating or preventing ocular hypertension and/or glaucoma, the compound comprising an amine-carboxy-monosulfonamide having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component, having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-disulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if administered topically in an aqueous eyedrop formulation to a mammalian eye.

140. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 139 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

141. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 139 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

142. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, an aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

143. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 142 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

144. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 142 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

145. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

146. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 145 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV; or, (b) lowering intraocular pressure upon topical ocular administration.

147. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 145 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

148. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, an aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least one sulfonamide component, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component, having at least one; C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

149. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 148 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

150. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 148 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

151. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, an aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure; and b. an amine-carboxy-alkyl component, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

152. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 151 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

153. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 151 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

154. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, an aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component other than diethylene-triamine-pentaacetate, the component having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

155. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 154 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inhibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV ; or, (b) lowering intraocular pressure upon topical ocular administration.

156. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 154 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.

157. A method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof, the method comprising administering ocularly to the subject, an aqueous eyedrop formulation comprising an amine-carboxy-sulfonamide compound or salt therof, the compound having:

a. at least two sulfonamide components, each component having at least one sulfonamide group bonded to a ring structure;

b. an amine-carboxy-alkyl component, having at least one C1-5 alkylamine component, at least one secondary or tertiary amine and at least one carboxy substituent, wherein the amine-carboxy-monosulfonamide compound. can have at least two isomeric forms and the compound is present in one substantially isomerically pure form, and wherein the amine-carboxy-sulfonamide compound has sufficient solubility in water at pH 7 to allow it to be dissolved at a concentration which can lower intraocular pressure if the formulation is administered topically to a mammalian eye.

158. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 157 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is more effective than a 2% (w/v) dorzolamide solution, as measured by either of the following activities: (a) inlibiting the enzymatic activity of at least one mammalian carbonic anhydrase enzyme selected from the group consisting of CA-II and CA-IV; or, (b) lowering intraocular pressure upon topical ocular administration.

159. The method of treating or preventing ocular hypertension and/or glaucoma in a subject in need thereof of claim 157 wherein the compound or salt, when formulated in a 2% (w/v) aqueous eyedrop solution, is effective in reducing intraocular pressure in normotensive rabbits by at least 3 mm Hg for at least four hours after administration.