Treatment of neuropathic corneal pain using NGF

Abstract

The present invention relates to a method of treating neuropathic corneal pain (NCP) comprising the administration of nerve growth factor (NGF).

Description

[Technical Field]

[0001] The present invention relates to a method for treating neuropathic corneal pain (NCP), comprising the administration of nerve growth factor (NGF). [Background technology]

[0002] Nerve growth factor (NGF) is a member of an evolutionarily well-conserved family of neurotrophin growth factors, including brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT3), and NT4 / 5, which are required for the development and survival of specific neuronal populations. NGF sequences are well-conserved across different species, with 90% homology between mouse and human NGF.

[0003] Recombinant human nerve growth factor (rhNGF) is approved by the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for use in the treatment of neurotrophic keratitis, a rare degenerative eye disease of the cornea characterized by decreased or lost corneal sensitivity, which may be asymptomatic or present with conjunctival hyperemia and slight vision loss during the early stages of the disease, ultimately leading to blindness. Recombinant human nerve growth factor (rhNGF) has also been shown to be safe and effective in improving the symptoms and signs of dry eye disease (DED) in human clinical trials (Sacchetti, et al., Br.J.Ophthalmol., 2020, 104, 127-135 (Reference 1)).

[0004] Dry eye disease (DED) is a multifactorial disorder of the ocular surface characterized by loss of tear film homeostasis, accompanied by ocular symptoms in which tear film instability and hyperosmolarity, inflammation and damage of the ocular surface, and sensory nerve abnormalities play a pathological role (Craig, et al., Ocul Surf., 2017, 15, 276-283 (reference 2)).

[0005] Neuropathic corneal pain (NCP), also known as neuropathic ocular pain, corneal neuralgia, keratoneuralgia, neuropathic-like pain, and chronic ocular surface pain, is caused by a complex interaction of various central and peripheral mechanisms, resulting in sensations such as burning, stabbing, eye pain, and general pain (Goyal and Hamrah, Seminars in Ophthalmology, 2016, 31, 59-70 (Reference 3)).

[0006] The International Association for the Study of Pain defines neuropathic pain as "pain that begins or arises from a primary lesion or dysfunction of the nervous system." Therefore, a diagnosis of neuropathic pain requires confirmation of injury or disease affecting the somatosensory pathways of the peripheral and / or central nervous system (CNS).

[0007] In patients with non-corporeal pain syndrome (NCP), the proparacaine challenge test is used to differentiate between peripheral and central pain. Treatment with 0.5% proparacaine hydrochloride eye drops reduces peripheral pain but not central pain (Goyal and Hamrah).

[0008] More recently, the following subtypes of NCP have been identified: patients may respond completely (peripheral NCP; approximately 25%) or partially (mixed NCP; approximately 50%) to anesthetic eye drops, or patients may not respond to anesthetic eye drops or their response may worsen (unpublished data).

[0009] In patients suffering from eye discomfort / pain, this discomfort / pain can be characterized as one of three types: nociceptive pain, inflammatory pain, and neuropathic pain. These different types of pain may also coexist.

[0010] Inflammation can be detected clinically (by redness on slit-lamp biomicroscopy); however, in many cases, inflammation can only be detected semi-clinically (for example, by the presence of dendritic cells by in vivo confocal microscopy).

[0011] The dendritic cell (DC) concentration in normal subjects is 25.9 ± 3.9 cells / mm³. 2 This is because the DC density is two standard deviations higher than normal (i.e., 75 cells / mm³). 2 (Super) indicates the presence of severe inflammation.

[0012] Recent data show that in typical dry eye disease (n=300 eyes), the DC density is 93.4±6.3 (Aggarwal, et al., Ocul Surf., 2021, 19, 183-189 (reference 4)).

[0013] Patients with neuropathic corneal pain (NCP) have a mean DC density of 71.89 ± 16.91 (Moein, et al., Ocul Surf., 2020, 18, 651-656 (Reference 5)).

[0014] Inflammation levels in NCP patients can vary considerably, ranging from no inflammation present, mildly elevated (less than 2 standard deviations above normal levels), or significantly elevated (more than 2 standard deviations above normal levels). [Prior art documents] [Non-patent literature]

[0015] [Non-Patent Document 1] Sacchetti,et al.,Br.J.Ophthalmol.,2020,104,127-135 [Non-Patent Document 2] Craig,et al.,Ocul Surf.,2017,15,276-283 [Non-Patent Document 3] Goyal and Hamrah, Seminars in Ophthalmology, 2016, 31, 59 - 70 [[Non - Patent Document 4]] Aggarwal, et al., Ocul Surf., 2021, 19, 183 - 189 [[Non - Patent Document 5]] Moein, et al., Ocul Surf., 2020, 18, 651 - 656 [[Summary of the Invention]]

[0016] The inventors have found that NGF is effective in treating neuropathic corneal pain (NCP) in patients suffering from NCP who respond to treatment with anesthetic eye drops. Furthermore, this result was confirmed in a mouse model of NCP that does not induce dry eye disease (DED).

[0017] [[ID=I8]] Therefore, the present invention relates to a method of treating neuropathic corneal pain (NCP) in a subject that requires treatment of neuropathic corneal pain (NCP), which comprises administering a composition comprising NGF.

[0018] The present invention also relates to NGF for use in treating neuropathic corneal pain (NCP) in a subject that requires treatment of neuropathic corneal pain (NCP).

[0019] A further aspect of the present invention relates to such a method comprising administering a composition comprising NGF, preferably rhNGF.

[0020] A further aspect of the present invention relates to NGF for use as defined above, wherein the NGF is rhNGF.

[0021] A further aspect of the present invention relates to NGF for a method or use as defined above, wherein the subject is a patient suffering from NCP and the patient is responsive to topical anesthesia treatment.

[0022] A further aspect of the invention relates to NGF for a method or use as defined above, wherein the subject is a patient suffering from peripheral or mixed NCP.

[0023] A further aspect of the invention relates to NGF for a method or use as defined above, wherein the subject is a patient suffering from NCP with mild inflammation.

[0024] A further aspect of the invention relates to NGF for a method or use as defined above, wherein the patient has a dendritic cell density (cells / mm 2 ) of less than 75, preferably less than 70.

[0025] A further aspect of the invention relates to NGF for a method or use as defined above, wherein the patient does not have dry eye disease (DED).

[0026] A further aspect of the invention relates to NGF for a method or use as defined above, wherein the patient does not have neurotrophic keratitis (NK).

[0027] A further aspect of the invention relates to NGF for a method or use as defined above, wherein a composition comprising NGF is administered topically to the ocular surface of the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] [Figure 1] Figure 1 shows the response to [5M] saline challenge at 3 days post-surgery (dps) for a mouse model of NCP, demonstrating an enhanced response in animals that received ciliary nerve ligation compared to sham treatment. [Figure 2]Figure 2 shows corneal fluorescein staining according to the NEI scale (0-15) for the mouse model of NCP, with higher scores indicating worsening defects in the corneal epithelium. The data demonstrate that there was no significant difference in CFS between the groups at any time point (p>0.05), which, importantly, indicates that the ligation model induced a pure NCP phenotype and not a dry eye disease phenotype. [Figure 3] Figure 3 shows the corneal sensitivity in a mouse model of NCP, measured by the Cochet-Bonnet sensory metric (an objective method for determining corneal nerve sensitivity by using monofilaments of different lengths and thereby applying different pressures to the cornea). [Figure 4] Figure 4 shows the response to the [5M] saline challenge throughout the entire course of the study in the mouse model of NCP. At 7, 10, and 14 dps, an observable decrease in the response to [5M] saline is present in the NGF-treated groups for both doses (p<0.05). By 14 dps, the NGF 4X / day group approaches recovery to the baseline response observed before NCP induction, and the NGF 6X / day group fully recovers to baseline. [Figure 5] Figure 5 shows the response to the chilled saline challenge in a mouse model of NCP. Responses varied, with significant differences observed between the NGF 4X / day group and the vehicle group on days 7, 10, and 14. [Figure 6] Figure 6 shows the responses to the L-menthol challenge in the mouse model of NCP. These responses were also varied. There was a significant difference between the groups at the initial stage, but this disappeared by day 14.

[0029] Detailed explanation This invention is based on the finding that NGF is effective in treating neuropathic corneal pain (NCP) in a specific and well-identified subset of patients suffering from NCP.

[0030] In particular, NGF is effective in treating patients with NCP who respond to treatment with topical anesthetics (e.g., propalacine) and have mild inflammation as assessed by IVCM. The use of NGF in this patient population results in improvements in pain levels and inflammation.

[0031] Accordingly, an object of the present invention is a method for treating neuropathic corneal pain (NCP) in subjects requiring treatment of NGF, comprising administering a composition containing NGF.

[0032] A further object of the present invention is NGF for use in the treatment of neuropathic corneal pain (NCP) in subjects requiring treatment of NCP.

[0033] The aforementioned subjects belong to the patient population for which the NGF treatment is effective, as described below.

[0034] According to the present invention, the patient population for which NGF treatment is effective can be defined as patients suffering from NCP who respond to treatment with a local anesthetic, preferably propalacine, and / or have mild ocular inflammation.

[0035] The expression "responding to treatment with a local anesthetic" in the present invention refers to partial or complete relief of pain as a result of local administration of an anesthetic eye drop for local use to the patient's eye, preferably the anesthetic being propalacaine, more preferably propalacaine hydrochloride, and more preferably propalacaine hydrochloride in a 0.5% dose.

[0036] Preferably, the patient having mild inflammation has a dendritic cell density (cells / mm²) of less than 75, preferably less than 70. 2 The patient has the following condition:

[0037] The patient population described above can be further defined as patients suffering from peripheral NCP or mixed NCP (i.e., patients who respond completely to anesthetic eye drops with complete pain relief, or partially with partial pain relief, respectively).

[0038] In addition to the above features, preferably, the patient population according to the present invention can be further defined as patients who do not suffer from dry eye disease (DED).

[0039] In addition to the above characteristics, preferably, the patient population according to the present invention can be further defined as patients who do not suffer from neurotrophic keratitis (NK).

[0040] As used herein, nerve growth factor (NGF) includes mouse NGF (mNGF) and recombinant human nerve growth factor (rhNGF) (containing senegermin-bkbj).

[0041] Preferably, the NGF is rhNGF.

[0042] The rhNGF for use according to the present invention can be produced in Escherichia coli (E. coli) using an expression vector incorporating the sequence of the proNGF mutant SP174-101 (SEQ ID NO: 5 in WO2013 / 092776), for example, according to the methods described in WO2000 / 022119 and WO2013 / 092776.

[0043] Preferably, according to the present invention, NGF is administered in the form of a pharmaceutical composition for use in the eyes.

[0044] Preferably, the NGF is administered topically to the surface of the eye of the target.

[0045] The precise dosage and regimen for administering NGF in the treatment or prevention of the above-mentioned conditions depends on many factors, such as the route of administration and the severity of the disease in the individual receiving treatment.

[0046] The pharmaceutical composition comprises the above-mentioned NGF and one or more ophthalmologically acceptable excipients.

[0047] An "ophthalmologically acceptable excipient" is an inactive excipient that allows for the delivery of a drug to the eye and / or eyelid to treat an eye disease or condition without adverse effects on the eye.

[0048] According to one embodiment, the ophthalmic composition may be a liquid eye drop composition for topical administration to the anterior segment of the eye.

[0049] The liquid composition may be in the form of a solution, emulsion, or suspension. The liquid composition may contain micelles. In one embodiment, the liquid composition is an aqueous composition.

[0050] Preferably, the liquid composition is an aqueous eye drop composition.

[0051] According to an alternative preferred embodiment, the ophthalmic formulation is a semi-solid ophthalmic formulation, preferably a cream, ointment, or gel.

[0052] In yet another preferred embodiment, the ophthalmic formulation is a solid ophthalmic formulation for the immediate preparation of the liquid or semi-solid ophthalmic formulation described above by adding a diluent before administration. Preferably, the solid ophthalmic formulation is in the form of a powder, more preferably in the form of a lyophilized powder.

[0053] Preferably, the liquid or semi-solid composition is ophthalmologically acceptable It contains ophthalmologically acceptable excipients selected from viscosity enhancers, penetration enhancers, buffers, osmolarity regulators, preservatives, and surfactants.

[0054] The thickening agent has the function of increasing the viscosity of the composition and improving its retention in the conjunctival sac, and is preferably selected from cellulose derivatives, preferably hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose; polyvinylpyrrolidone; and a gelling agent, preferably guerlain, xanthan gum, and carbopol-974.

[0055] The permeability enhancer has the function of increasing drug permeability across the ocular membrane and is preferably selected from cyclodextrin, chelating agents, crown ethers, bile acids, and bile salts.

[0056] Buffers have the function of providing and maintaining the correct pH of the formulation, preferably between 6 and 8, to make it suitable for ophthalmic use. Preferred buffers are phosphate buffers, but other buffers that can maintain pH within the desired range, particularly those suitable for ophthalmic use, are also included.

[0057] An osmotic regulator is a salt that can make a liquid composition isotonic with ophthalmic solution. A preferred salt is sodium chloride (NaCl), but other bioacceptable salts, such as potassium chloride (KCl), calcium chloride (CaCl2), and magnesium chloride (MgCl2), or mixtures thereof, may also be used.

[0058] Preservatives inhibit microbial activity. Suitable preservatives include quaternary ammonium compounds, such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0059] The surfactant has the function of stabilizing the composition and reducing or preventing NGF adsorption to various surfaces of the container, and is preferably selected from polysorbates such as Tweeny80, poloxamers such as PluronicsF68, or proteins such as serum albumin.

[0060] The aforementioned liquid eye drop composition may be part of a kit comprising the composition, a container for containing the composition, and a droplet dispenser.

[0061] The NGF aqueous composition according to the present invention may contain a sufficient amount of bioacceptable salts to provide the correct fluid tonicity and maintain NGF in solution.

[0062] Other additives commonly used in pharmaceutical aqueous compositions and known to those skilled in the art, such as sugars, sugar alcohols, amino acids, cellulose derivatives, and polyethylene glycol, may be present in the NGF aqueous composition.

[0063] The NGF aqueous composition contains a sufficient amount of water to achieve the appropriate concentration of the compositional components.

[0064] The liquid composition contains NGF at a therapeutically effective concentration.

[0065] Preferably, in the liquid composition, NGF is present at a concentration in the range of about 0.0001% to about 0.5% w / v of the aqueous composition, more preferably about 0.001% to about 0.1% w / v, and most preferably about 0.002% w / v.

[0066] Preferably, the NGF is administered 3 to 6 times a day for a period of 4 to 12 weeks. The administration schedule is determined by a physician based on the dosage of the NGF preparation and the severity of the patient's condition.

[0067] Some aspects of the present invention will be more successfully described below:

[0068] In the first embodiment, an NGF is provided for use in the treatment of neuropathic corneal pain (NCP) in subjects requiring treatment of NCP.

[0069] In a second embodiment, in the preceding embodiment, the NGF for use is selected from mouse NGF (mNGF) and recombinant human nerve growth factor (rhNGF), preferably recombinant human nerve growth factor (rhNGF).

[0070] In a third aspect, in any of the preceding aspects, the subject is a patient suffering from neuropathic corneal pain (NCP), responding to treatment with a local anesthetic, and / or having mild ocular inflammation.

[0071] In a fourth embodiment, in the third embodiment, the subject is a patient having partial or complete relief of pain as a result of local administration of an anesthetic eye drop for local use to the eye.

[0072] In the fifth aspect, in either the third or fourth aspect, the anesthetic is propalacaine, more preferably propalacaine hydrochloride, more preferably propalacaine hydrochloride in a 0.5% dose. In the sixth embodiment, in any of the preceding embodiments, the patient has a dendritic cell density (cells / mm²) of less than 75, preferably less than 70. 2 ) has.

[0073] In the seventh embodiment, in any of the preceding embodiments, the subject does not suffer from dry eye disease (DED).

[0074] In the eighth aspect, in any of the preceding aspects, the subject is not suffering from neurotrophic keratitis (NK).

[0075] In the ninth embodiment, the NGF for use described above is administered topically to the surface of the eye of the subject in any of the preceding embodiments.

[0076] In the tenth embodiment, in any of the preceding embodiments, the NGF is administered in the form of a pharmaceutical composition for use in the eye.

[0077] In the eleventh aspect, a pharmaceutical composition is provided for use in the treatment of neuropathic corneal pain (NCP) in subjects requiring treatment of neuropathic corneal pain (NCP) as defined in any of the second to eighth aspects.

[0078] In the twelfth embodiment, in either the tenth or eleventh embodiment, the pharmaceutical composition is a liquid formulation composition for topical administration to the anterior segment of the eye.

[0079] In the 13th embodiment, in the 12th embodiment, the liquid composition is an aqueous composition, preferably an aqueous eye drop composition.

[0080] In the fourteenth embodiment, in either the tenth or eleventh embodiment, the pharmaceutical composition is a semi-solid ophthalmic preparation, preferably a cream, ointment, or gel.

[0081] In the 15th aspect, in either the 10th or 11th aspect, the pharmaceutical composition is a solid ophthalmic formulation for the immediate preparation of a liquid or semi-solid ophthalmic formulation according to the 12th or 14th aspect. [Examples]

[0082] example The following examples illustrate the present invention without limiting its scope.

[0083] Example 1 - A retrospective case study of a patient with neuropathic corneal pain (NCP) A retrospective case series was conducted between December 2018 and January 2021 of 20 patients with neuropathic corneal pain who visited the Corneal Department of the New England Eye Center (Tufts Center for Therapeutic Research, Boston, MA). Patients were diagnosed with NCP if symptoms were disproportionate to signs, if the patient's pain did not respond to extensive prior treatment, if symptoms did not subside after 90 seconds with anesthetic eye drops, or if the patient showed microneuromas in the IVCM. In cases where pain completely subsided with anesthetic eye drops, lack of pain response to extensive prior treatment, symptoms disproportionate to signs, decreased corneal sensation, and the presence of microneuromas were used to diagnose NCP. Recombinant nerve growth factor (Oxervate (商標) Prior to the procedure, a thorough medical record review was conducted, and clinical parameters were recorded at two time points. The patient had rhNGF (Oxervate (商標) Patients were treated with rhNGF (6x / day for 8 weeks). After initiating rhNGF treatment, patients were followed up, pain severity reported by the patients was recorded, and IVCM (intravenous pulmonary difficulty) was administered before and after the completion of the 8-week treatment period.

[0084] This study was approved by the Institutional Board Review of Tufts Medical Centre / Tufts University Health Sciences. The protocol conforms to the Declaration of Helsinki and the Health Insurance Portability and Accountability Act (HIPAA).

[0085] Data extracted from medical records included patient demographics, clinical findings (including vital pigment staining using the Oxford scale and tear metrics), symptom questionnaires (including the Ocular Surface Disease Index (OSDI)), and ocular pain assessment surveys (OPAS). Furthermore, results from propalacaine challenge trials (PCTs) from the same visit were recorded. For PCTs, patients were asked to report their pain relief based on a visual analog scale 90 seconds after instillation of 0.5% propalacaine hydrochloride eye drops (Alcaine; Novartis Ophthalmics, East Hanover, NJ).

[0086] Laser IVCM (Heidelberg Retina Tomograph 3 / Rostock Cornea Module, Heidelberg Engineering GmbH, Heidelberg, Germany) was routinely performed on all patients presenting with pain and discomfort to confirm subbasal corneal nerve changes, as previously described (Cruzat, et al., IVOS, 2011, 52, 5136-5143). IVCM was obtained from all patients and compared to age- and sex-matched reference controls from a prospectively registered healthy human database. Images were obtained using a 63x objective immersion lens with a numerical aperture of 0.9 (Olympus, Tokyo, Japan). This microscope uses a 670 nm red wavelength diode laser source that produces images representing a 400 × 400 μm (horizontal × vertical) coronal section of the cornea. Digital images were recorded at 30 frames / second. Adjacent images are separated by only 1 μm with an azimuthal resolution of 1 μm / pixel. To perform this procedure, both eyes were locally anesthetized with 0.5% propalacaine hydrochloride (Alcaine; Novartis Ophthalmics). Subsequently, one drop of 2.5% hydroxypropyl methylcellulose (GenTeal gel, Alcon, Fort Worth, TX) was administered to improve optical coupling with the corneal module of the microscope. A disposable sterile polymethyl methacrylate cap (Tomo-Cap; Heidelberg Engineering GmbH) filled with a layer of 2.5% hydroxypropyl methylcellulose (GenTeal gel; Alcon) was placed on the corneal module, and the gel was also applied to the surface of the cap. The apparatus was manually advanced until the gel on the cap contacted the surface of the central part of the cornea. 50–100 images of the subbasal layer of the cornea were obtained in 6–8 sequences, and a masked observer (PH) selected the three most representative images determined to have the best focus, single layer, minimal overlap, and good contrast of the subbasal plexus. One anonymous observer (PH) analyzed IVCM images for dendritic cell (DC) density.

[0087] As can be seen from the data shown in Table 1, patients who reacted to proparacaine and had mild inflammation as observed by IVCM responded to Oxervate (商標) treatment and showed significant improvement (i.e., a decrease in pain level). Patients who did not respond to proparacaine or had severe inflammation did not show a decrease in pain level in response to Oxervate (商標) treatment. Furthermore, patients without inflammation, as well as those with mild or moderate inflammation, also showed improvement regarding inflammation.

[0088] [Table 1]

[0089] Example 2 - Evaluation of topical NGF in a mouse model of neuropathic corneal pain This study was conducted to investigate the efficacy of topical NGF eye drops (0.02 mg / mL) in the treatment of neuropathic corneal pain (NCP) using a mouse model of NCP induced by ciliary nerve ligation.

[0090] Confirmation of ciliary nerve ligation and induction of NCP. [5M] Baseline responses to saline were obtained before ligation or sham surgery. On the third day post-surgery (dps), the animals were challenged again with [5M] saline. As shown in the results of Figure 1, the ligation was successful as the animals that received the ligation treatment had enhanced responses (indicating an increase in pain) (p < 0.0001), as determined by [5M] saline challenge.

[0091] Test design On day 3, the ligated animals were divided into two groups – one group received recombinant NGF (0.02 mg / mL), and the other group received vehicle eye drops six times per day. The mice were followed up to 14 dps, with outcome measurements including corneal fluorescein staining, Cochet-Bonnet sensory measurements, [5M] saline challenge, chilled saline challenge, and L-menthol challenge performed at 7, 10, and 14 dps. At 14 dps, the cornea and trigeminal ganglion were collected for further analysis.

[0092] Clinical outcomes Corneal fluorescein staining: Mice were briefly anesthetized with ketamine / xylazine and prepared for examination of the ocular surface using slit-lamp microscopy. One drop of fluorescein was instilled into the ligated eye, and the mice were manually instructed to blink to cover the entire ocular surface. The stain was allowed to develop for at least one minute, at which point the ocular surface was evaluated for staining and scored using the National Eye Institute scale (0-15). This is a necessary quality control measure to ensure that the ligated animals do not develop dry eye disease (NEI score > 10) (which may confuse the results). As shown in Figure 2, only slight staining was present in the animals across all groups and time points, indicating that the ligation approach is inducing a "pure" neuropathic corneal pain phenotype rather than dry eye disease. Furthermore, these staining scores are consistent with what would be expected from naive mice (slight staining).

[0093] Corneal sensation: To confirm that corneal sensation is intact in all animals, a Cochet-Bonnet sensory meter was used. This approach uses a thin nylon filament applied to the central part of the cornea. Sufficient pressure is applied to cause a slight bend in the filament. Based on the length of the filament, it is possible to determine the amount of pressure applied. If corneal sensation is intact, the outcome is the blink reflex in the animal.

[0094] For these assessments, individual mice had to exhibit a positive response (blinking) in 2 / 3 of the attempts; otherwise, the filament length was reduced in 5 mm increments for reassessment (shorter filament lengths indicated greater pressure). As shown in Figure 3, there were slight differences in corneal nerve sensation between the NGF 4X / day group and the vehicle group on days 7 and 10 (p<0.05 and p<0.01, respectively), but not on day 14, the most important time point (p>0.05). These results were analyzed by repeated measures ANOVA.

[0095] However, importantly, corneal sensation was essentially intact at a similar level across all groups. This indicates that ligation does not destroy the ciliary nerve (if it did, the blink reflex would be completely absent). Instead, the nerve is preserved but functionally impaired.

[0096] Response to hyperosmolar saline: The following were confirmed: (1) ligation was successful, (2) ligation did not cause dry eye disease as evaluated by corneal fluorescein staining, and (3) corneal sensation remained intact despite ligation. Changes in pain outcomes were then investigated. Hyperosmolar saline solution [5M] was instilled into the ocular surface on the ligated side (10 μL), and the number of paw wipes was counted within a 30-second interval immediately afterward. The number of paw wipes was used as a measure of pain, with a higher number of paw wipes indicating greater pain experienced by the animal. The results are shown in Figure 4.

[0097] As shown in Figure 4, similar baseline responses were observed between the groups on day 0 and on day 3 post-ligation, when tissue was collected before the treatment began. During the follow-up period, both the 6X and 4X / day doses resulted in a significant reduction in limb wiping (and therefore pain) in the NGF treatment groups. Most notably, the 6X / day NGF group showed a recovery to the baseline response. Furthermore, the vehicle treatment group showed no change in response.

[0098] Response to cold saline: Cold saline was instilled into the eye surface on the ligated side (10 μL), and the number of times the limb was wiped within a 30-second interval immediately afterward was counted. The number of times the limb was wiped was used as a measure of pain, with a higher number of wipes indicating greater pain experienced by the animal. The results of this assessment are shown in Figure 5. As shown there, there was a statistically significant difference between the NGF 4X / day group and the vehicle group, but this was not observed in the NGF 6X / day group. This result requires further research to determine its biological significance and is likely a result of different dependencies between polymodal nociceptors and cold nociceptors on NGF.

[0099] Response to L-menthol: L-menthol can activate M member 8 (TRPM8), a transient receptor potential cation channel subfamily that plays a major role in cold sensation. Therefore, L-menthol was instilled into the eye surface on the ligated side (10 μL), and the number of times the limb was wiped within a 30-second interval immediately afterward was counted. The number of limb wipes was used as a measure of pain, with a higher number of wipes indicating greater pain experienced by the animal. The results of this assessment are shown in Figure 6. As shown there, there was a statistically significant difference between the NGF 4X / day group and the vehicle group, but this was not observed in the NGF 6X / day group. Interestingly, these results are similar to those observed with cold saline alone. These results require further research to determine their biological significance and are likely a result of different dependencies between polymodal nociceptors and cold nociceptors on NGF.

[0100] On day 14, the trigeminal ganglion (TG) was resected for qRT-PCR analysis of neurotrophic factors and cytokines.

[0101] NGF treatment reduced levels of several neurotrophic factors in TG compared to vehicle treatment (BDNF: 0.78 vs. 1.00, p<0.05; NT-3: 0.25 vs. 1.00, p<0.01; NT-4 / 5: 0.11 vs. 1.00, p<0.05), but did not show an increase in pro-inflammatory cytokines (IL-1b: 0.50 vs. 1.00, IL-6: 0.92 vs. 1.00, TNF-a: 0.69 vs. 1.00; all p>0.05).

[0102] Summary of clinical outcomes: These data suggest that NGF eye drops have a beneficial effect at both the tested doses (6x / day and 4x / day). Importantly, there were no differences in corneal fluorescein staining between the groups at any time point, and therefore this model does not induce dry eye disease.

[0103] As assessed by Cochet-Bonnet sensory measurements, there was a slight difference in corneal sensitivity at the initial time, but this disappeared by day 14. Most importantly, the ligation did not destroy the ciliary nerve, as corneal sensitivity remained intact. Importantly, there was a very significant improvement in response to the 5M saline challenge, which was the most useful pain outcome. This effect was observed at all time points in both the 6X and 4X NGF-treated groups, with the 6X group returning completely to baseline response by day 14. Outcomes with hypothermia and L-menthol were more varied and may indicate either an initial benefit of NGF, differences in nociceptor dependence to NGF, or a result of improvements in surgical technique over time.

[0104] In summary, data from retrospective clinical trials of patients with NCP, and data from mouse models of NCP, demonstrate that treatment with rhNGF results in an improved pain response in a subset of patients with NCP. The data further demonstrate that NGF treatment results in an improvement in inflammation in a subset of patients with NCP. * * * * *

[0105] The present invention is not limited to the scope of the specific embodiments described herein. In fact, various modifications in addition to the embodiments described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

[0106] All patents, applications, publications, test methods, documents, and other materials cited herein are incorporated herein by reference. While this application relates to the invention described in the claims, it may also encompass the following other embodiments. 1. A method for treating neuropathic corneal pain (NCP) in subjects requiring treatment of NGF, comprising administration of a composition containing NGF. 2. NGF for use in the treatment of neuropathic corneal pain (NCP) in patients requiring treatment for NCP. 3. The method described in 1 above or the NGF for use described in 2 above, wherein the NGF is rhNGF. 4. The method or use of NGF according to any one of 1 to 3 above, wherein the subject is a patient suffering from NCP who responds to treatment with a local anesthetic, preferably propalacaine, and / or has mild inflammation. 5. The subject has a dendritic cell density (cells / mm²) of less than 75, preferably less than 70. 2 NGF for use according to any one of the above 1 to 4, having ) 6. The method or use of NGF according to any one of items 1 to 5 above, wherein the subject is a patient suffering from peripheral or mixed type NCP. 7. The method or use of NGF according to any one of items 1 to 6 above, wherein the subject is a patient who does not suffer from dry eye disease (DED) and / or neurotrophic keratitis (NK). 8. The method or use of NGF according to any one of 1 to 7 above, wherein NGF is administered topically to the ocular surface of the subject in the form of a pharmaceutical composition for use in the eye. 9. The method or use of NGF according to 8 above, wherein NGF is present in the pharmaceutical composition at a concentration in the range of about 0.0001% to about 0.5% w / v, more preferably about 0.001% to about 0.1% w / v, and most preferably about 0.002% w / v. 10. The method or use of NGF according to any one of the above 1 to 9, wherein the NGF is administered between 3 and 6 times a day for a period of 4 to 12 weeks.

Claims

1. A pharmaceutical composition for treating neuropathic corneal pain (NCP) in subjects requiring treatment of NCP, comprising a therapeutically effective concentration of NGF, It is formulated for administration to the ocular surface of the aforementioned target, NGF is present in the pharmaceutical composition at a concentration in the range of approximately 0.0001% to approximately 0.5% w / v. The aforementioned pharmaceutical composition.

2. The pharmaceutical composition according to claim 1, wherein the NGF is rhNGF.

3. The pharmaceutical composition according to claim 1 or 2, wherein the subject is a patient suffering from NCP who responds to treatment with a local anesthetic and / or has mild inflammation.

4. The aforementioned subjects have a dendritic cell density (cells / mm²) of less than 75. ２ A pharmaceutical composition according to claim 1 or 2, having the following characteristics:

5. The pharmaceutical composition according to claim 1 or 2, wherein the subject is a patient suffering from peripheral or mixed type NCP.

6. The pharmaceutical composition according to claim 1 or 2, wherein the subject is a patient who does not suffer from dry eye disease (DED) and / or neurotrophic keratitis (NK).

7. The pharmaceutical composition according to claim 1 or 2, packaged for administration of NGF three to six times a day for a period of four to twelve weeks.

8. The pharmaceutical composition according to claim 3, wherein the local anesthetic is propalacaine.

9. The aforementioned subjects have a dendritic cell density (cells / mm²) of less than 70. ２ The pharmaceutical composition according to claim 4, having )

10. A pharmaceutical composition according to claim 1 or 2, wherein NGF is present in the pharmaceutical composition at a concentration in the range of about 0.001% to about 0.1% w / v.

11. A pharmaceutical composition according to claim 1 or 2, wherein NGF is present in the pharmaceutical composition at a concentration of about 0.002% w / v.

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