DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Acknowledgement of Receipt
Applicant’s Response, filed 6/5/2025, in reply to the Office Action mailed 3/5/2025, is acknowledged and has been entered. Claims 1 and 13 have been amended. Claims 1, 3, 6-9 and 11-14 are pending and are examined herein on the merits for patentability.
Response to Arguments
Applicant’s arguments have been fully considered. The previous rejections have been modified in view of claim amendment. The Examiner’s response to Applicant’s arguments is incorporated below.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claim(s) 1, 3, 5-9, 11, 12 and 14 are rejected under 35 U.S.C. 103 as being unpatentable over Liao et al. (US 2015/0056273, hereinafter Liao I), in view of Liao et al. (2012 IEEE International Ultrasonics Symposium Proceedings, p. 440-443, hereinafter Liao II), in further view of Noda et al. (Ultrasound in Medicine & Biology, 2013, 39(3), p. 413-423) and Shih et al. (J. Controlled Release, 2013, p. 167-174).
Liao I teaches an external type microbubble ultrasound contrast agent may employ a medium, either aqueous or a gel form, and contain microbubbles of a specific particle size and at a specific concentration. The material of the microbubbles may be albumin, liposomes, polymers, copolymers or mixtures of the aforementioned material or a combination of those above. The material of the microbubbles may also include pentose and/or hexose. A method for preparing an external type microbubbles ultrasound contrast agent of topical uses is also provided. The microbubble ultrasound contrast agent may have microbubbles with different particle sizes by adjusting the percentage of the medium and the material of the microbubbles the mixed solution and followed by the same ultrasonic oscillating steps which oscillating the mixed solution for about 100 to about 140 seconds. The external type microbubble ultrasound contrast agent may be applied in conjunction with the application of mechanical oscillation waves. Through a series of swelling and shrinking processes induced by the oscillation energy of the mechanical oscillation waves, the microbubbles burst or destructed and the generated energy and shock waves lead to minor damages of cells or tissues, which further strengthen the absorption of applied chemicals or small molecules, also the microbubbles with different particle sizes may lead to different penetration depth for the applied chemicals or small molecules. The commonly used energy source of the mechanical oscillation waves may be a source of an optical energy or acoustic energy, such as an ultrasound source or a laser source. The external type microbubble ultrasound contrast agent of the present invention, suitable for applying onto a local region of the body surface of the living body, may be used in combination with the mechanical wave(s) generated by the mechanical oscillating energy source to cause the microbubbles in the external type microbubble ultrasound contrast agent bursting to produce energy and shock waves. The energy and the shock waves from microbubble bursting cause minor and reversible damages on the contact area of the skin surface or mucous membrane, thereby increasing the percutaneous absorption of chemicals or small molecules. The microbubble ultrasound contrast agent may be widely used in medical or beauty fields, to help strengthen the absorption of painkillers after surgery or the absorption of beauty care ingredients (paragraph 0006).
The microbubble contrast agent can enhance the delivery of the chemical and promote the absorption of the drug or small molecules (paragraph 0103).
The preparation of the microbubble contrast agent is set forth in paragraphs 0062+, including providing an isotonic saline solution comprising microbubbles, (Component A, the microbble liquid); a drug or other chemical to be delivered (Component B) and Component B is used a diluent to dilute Component A 2-1000 times and the composition obtained after dilution is applied to the surface of the body.
The concentration of the particles is preferably 2x106 – 2x108 particles/ml (paragraph 0066).
The size of the microbubbles is 0.5 to 3.7 micrometers (paragraph 0059).
The material of the microbubbles is albumin, liposomes, polymers, etc. (paragraph 0059).
The external use microbubble contrast agent of the present invention may be used in the ear treatments. The microbubble contrast agent of this invention is mixed with the dye and/or one or more medical ingredients and administrated to the inner ear of guinea pigs. The administration of the mixtures may be conducted in different ways to test the delivery efficiency of the dye or the ingredient (paragraph 0099).
The animals used in the test are 60 guinea pigs with the normal Preyer's reflex to the sound(s) and are divided into three groups with the following experimental conditions: (1) the tympanic bullae of 24 guinea pigs are filled with the microbubble ultrasound contrast agent mixed the dye indicator and applied with the ultrasound (paragraph 0101).
Sonoporation Gene Transfection System (ST2000V, NepaGene, Japan), with a probe size of 6 mm and the waveform of square waves. In the experiments, the ultrasound is operated at a frequency of 1 MHz, a duty cycle of 50%, energy of 3 W/cm.sup.2, is applied for 1 minute. In the experiments, the probe is placed on the body surface facing the round window membrane with a distance of 5 mm (paragraph 0102).
FIG. 12 shows the results of the delivery efficiency using different administration approaches of the microbubble contrast agent in the inner ear treatment experiments.
Compared to the control group of delivering the dye or drug into the inner ear through the diffusion effect, the experimental results indicate that the ultrasound used together with the microbubble ultrasound contrast agent can enhance the drug delivery efficiency. In addition, in order to deliver gentamycin into the inner ear, the microbubbles ultrasound contrast agent of this invention is used along with the application of the ultrasound. By using such approach, the concentration of gentamycin delivered into the cochlear tissues is significantly higher than that of the control group without applying the ultrasound. Hence it is confirmed that the microbubble contrast agent can enhance the delivery of the chemical and promote the absorption of the drug or small molecules.
FIGS. 13A-13F show the delivery results of the green dye indicator entering into the round window membrane cells of the inner ear under different administration approaches. FIGS. 13A-136C show the delivery results of the experimental groups using the ultrasound microbubble contrast agent mixed with the green dye indicator and operated with the ultrasound. FIGS. 13D-13F show the delivery results of the control groups using the ultrasound microbubble contrast agent mixed with the green dye indicator but without applying the ultrasound (through the diffusion effect). Compared the results of little or no green dye entering into the round window membrane cells in FIGS. 13D-136F, the results of FIGS. 13A-13C show much more green dyes entering into the round window membrane cells (paragraph 0103).
FIGS. 13A-13F show the delivery results of the green dye indicator entering into the round window membrane cells of the inner ear under different administration approaches. FIGS. 13A-136C show the delivery results of the experimental groups using the ultrasound microbubble contrast agent mixed with the green dye indicator and operated with the ultrasound (paragraph 0104).
Liao II teaches that direct drug delivery into inner ear can be achieved by three approaches: (1) diffusion of drug through round window; (2) injection of drug through round window; (3) cochleostomy or canalostomy. The latter two approaches are invasive and have the risk of hearing loss and vertigo. Drug diffusion through round window is only the noninvasive approach. However, how to enhance drug diffusion and how to noninvasively promote drug delivery through round window are two issues that need to be investigated. In this study, we target on the practical application of microbubbles (MBs)-ultrasound on increasing the wound window membrane (RWM) permeability for facilitating drug or medication delivering into the inner ear. Using biotin-FITC conjugates (biotin-FITC) as delivered agents and performed on guinea pigs animal models, we showed that the MBs-ultrasound exposure can greatly improve the inner ear system utility of the biotin-FITC delivery via RWM at different kinds of approaches about 3.5 to 38 times compare to that solely soaking biotin-FITC around the RWM for spontaneous diffusion. In addition, significant enhancement of hair cells uptake of gentamicin was demonstrated in animals whose tympanic bullas were soaked with MBs-mixed gentamicin−Texas Red or gentamicin and exposed to ultrasound. Furthermore, the increased permeability of RWM resulted from acoustic cavitation of MBs could also be visualized immediately following ultrasound exposure by using Alexa Fluor 488-conjugated phalloidin as a tracer. Most importantly, such applications were shown without resulting damage to the integrity of RWM or deterioration of the hearing thresholds assessed by auditory brainstem responses, suggesting this MBs-ultrasound not only benefits in developing therapeutic strategies for inner ear diseases, but also help in providing a more precise and well-controlled release for medications passing through the RWM (abstract).
Albumin-shelled MBs were prepared and mixed with gentamicin-Texas red conjugate. A fenestration allows exposing the round window, loading the mixture of MBs, and delivering reagents into the middle ear cavity for sonication.
A fenestration (approximately 4 mm in diameter) was created in the tympanic bulla by drilling with diamond burrs. This fenestration allows exposing the round window, loading the mixture of MBs, and delivering reagents into the middle ear cavity for sonication (page 441).
It was shown that ultrasound-aided microbubbles promote the delivery of biotin to the inner ear. Cavitation of MBs-ultrasound results in increase of round window membrane permeability without causing complete cell damage. Microbubble-ultrasound exposure did not deteriorate hearing acuity. The MBs-ultrasound system appears to be safe in our guinea pigs model. Even though, a comprehensive histology study of the inner ear following MBs-ultrasound exposure is needed before final conclusions concerning safety issue can be made (page 442).
Accordingly, Liao I and II teach a method of delivering drugs to inner ear facilitated by microbubbles comprising providing a microbubble-drug mixture and applying to the middle ear cavity and sonication by ultrasonic waves to produce cavitation so as to increase permeability of the round window membrane allowing the drug in the microbubble-drug mixture to penetrate the round window membrane. However, Liao I recites that microbubbles are in direct contact with the ultrasound probe to induce cavitation under the ultrasound energy (paragraph 0057), rather than in indirect contact as claimed. Liao I and II do not specifically recite a mechanical oscillation wave source inserted in the external ear canal in direct contact with the second medium.
Noda teaches low-intensity focused ultrasound (LIFU) increases vessel permeability and antibacterial drug activity in the mouse middle ear. We determined appropriate settings by applying LIFU to mouse ears with the external auditory canal filled with normal saline and performed histologic and immunohistologic examination. Acute otitis media was induced in mice with nontypable Haemophilus influenzae, and they were given ampicillin (50, 10, or 2 mg/kg) intraperitoneally once daily for 3 days with or without LIFU (1.0 W/cm2, 20% duty cycle, 30 s). In the LIFU(+) groups receiving the 2- and 10-mg/kg doses, viable bacteria counts, number of inflammatory cells and IL-1β and TNF-α levels in middle ear effusion were significantly lower than in the LIFU(−) groups on the same doses. Severity of AOM also tended to be reduced more in the LIFU(+) groups than in the LIFU(−) groups. LIFU application with antibiotics may be effective for middle ear infection (page 413).
For example, the effects of LIFU on AOM was tested. In mice with MEE filling the middle ear, only the external auditory canal was filled with normal saline solution, and LIFU was applied through the TM. In this study, an incisional myringotomy was not performed because of the filling of the middle ear with MEE. This schedule of antibiotic administration has been described previously (page 415).
Our results show that in vivo LIFU can disrupt vessels and increase vessel permeability in the mouse middle ear mucosa without causing tissue damage. The first part of this study showed the maximum LIFU parameters that would not cause histologic damage in the murine middle ear mucosa: 1.0 W/cm2 with a duty cycle of 20% for 30 s. When ampicillin was given for 3 d at intraperitoneal doses of 2 and 10 mg/kg, the number of inflammatory cells, viable bacteria counts, and levels of IL-1b and TNF-a in MEEs were significantly lower when LIFU was administered to the ear than when it was not. LIFU application along with antibiotics may be an effective strategy for treatment of AOM (page 422-23).
Shih teaches that the round window membrane (RWM) acts as a barrier between the middle ear and cochlea and can serve as a crucial route for therapeutic medications entering the inner ear via middle ear applications. In this study, we targeted the practical application of microbubbles (MBs) ultrasound on increasing the RWM permeability for facilitating drug or medication delivery to the inner ear. Using biotin–fluorescein isothiocyanate conjugates (biotin–FITC) as delivery agents and guinea pig animal models, we showed that MB ultrasound exposure can improve the inner ear system use of biotin–FITC delivery via the RWM by approximately 3.5 to 38 times that of solely soaking biotin–FITC around the RWM for spontaneous diffusion. We also showed that there was significant enhancement of hair cell uptake of gentamicin in animals whose tympanic bullas were soaked with MB-mixed gentamicin–Texas Red or gentamicin and exposed to ultrasound. Furthermore, increased permeability of the RWM from acoustic cavitation of MBs could also be visualized immediately following ultrasound exposure by using Alexa Fluor 488-conjugated phalloidin as a tracer. Most importantly, such applications had no resulting damage to the integrity of the RWM or deterioration of the hearing thresholds assessed by auditory brainstem responses. We herein provide a basis for MB ultrasound-mediated techniques with therapeutic medication delivery to the inner ear for future application in humans (abstract).
Although intracochlear application can achieve greater bioavailability of drugs entering the inner ear than the intratympanic approach, the former usually requires invasive surgical manipulation to interrupt the inner ear structure, and may increase the risk of deafness. Currently in clinical practice, intracochlear injection is usually accompanied by cochlear prosthesis implantation. The intratympanic approaches, in contrast, enter the scala tympani and disperse within the inner ear, relying solely on diffusion through the RWM. To date, a wide range of materials and methods have been developed for enhancing local drug delivery to the inner ear via the RWM; these include hydrogels, nanoparticles, and devices for active intratympanic drug delivery (page 167-8).
Therapeutic ultrasound exposure can be provided using either nonfocused transducers (plane wave) or spherically focused transducers. High-intensity-focused ultrasound can focus the acoustic waves onto very small volumes, which greatly increases their intensity and allows energy to be deposited deep inside the body [15,16]. Nonfocused ultrasound is typically used for physical therapy applications and for enhancing transdermal delivery, for example, sonophoresis. Recently, ultrasound combined with a contrast agent, microbubbles (MBs), has been used to target or control drug release to tissues and cells.
The RWM can be a new target for evaluating the biological effects resulting from such nonfocused MB-ultrasound exposure and the efficiency in drug delivery. The purpose of this study was to determine whether ultrasound activated MBs can enhance local drug delivery to the inner ear through the RWM by examining laboratory-made albumin-shelled MBs in a guinea pig model (page 168).
Ultrasound (ST2000V, Nepagen) equipped with a 6-mm diameter transducer was used for irradiation in the experiments.
It would have been obvious to one of ordinary skill in the art at the time of the invention to provide an ultrasonic probe in the ear canal as a means to apply the mechanical oscillation wave source in the methods of Liao I and II when the teachings are taken in view of Noda. One would have been motivated to do so because Liao is concerned with non-invasive promotion of drug delivery through round window, including aversion to interrupting the inner ear structure (Liao II, page 440). One would have had a reasonable expectation of success in doing so because Noda teaches applying LIFU to mouse ears with the external auditory canal filled with normal saline and shows that LIFU can disrupt vessels and increase vessel permeability in the mouse middle ear mucosa without causing tissue damage and that LIFU application along with antibiotics may be an effective strategy for treatment of AOM (page 422-23). With regard to the limitation directed to a second solution in the external ear canal and wherein the mechanical oscillation wave source is in contact with the second solution in the external ear canal, saline is taught by Noda in the external auditory canal. With regard to claims 11-12, Liao I teaches that the concentration of the contrast agent may be in the range of 2x106-2x108 particles/ml, which encompasses the claimed ranges (paragraph 0060).
With regard to the limitation of “applying the microbubble-drug mixture onto a round window membrane of a middle ear cavity” it is noted that the round window one of the two openings from the middle ear into the inner ear and is sealed by the round window membrane. The instant specification recites that “the eardrum was punctured by a 22G needle and injected about 300 μL of a 10-fold diluted microbubble ultrasound contrast agent mixed with Biotin-FITC solution into the middle ear cavity (paragraph 0067). In Example 5, Liao 1 recites: “A 22G needle is used to puncture the eardrum, and 200 μL of the temperature-sensitive microbubbles drug (DEX) hydrogel of the present disclosure is applied into the middle ear cavity.” Accordingly, since both the instant specification and Liao 1 recite application via the same method it is considered that applying the microbubble-drug mixture onto a round window membrane of a middle ear cavity is achieved.
With regard to the amended claim limitation “wherein a 3 W/cm2 ultrasound is applied to the skull skin behind the ear corresponding to the tympanic bulla 3 times and 1 minute each time, and the microbubble-drug mixture in the middle ear cavity is replaced every time to ensure that the microbubble-drug mixture produces the cavitation each time,” see page 442 of Liao II. To determine the efficiency of MBs-ultrasound-mediated delivery in the inner ear system, biotin-FITC conjugates (biotin-FITC) were used as delivered agent. Animals were divided into several experimental groups. In ultrasonic microbubble (USM) group, tympanic bulla was filled with 200 μ1 mixture of biotin-FITC (40 ug/ml) and MBs, while in ultrasonic (US) group only 200 μ1 biotin-FITC solution (40 ug/ml) was filled. Ultrasound exposures were then applied to both groups for 1 min. In USM-x2 and US-x2 groups, the same irradiation procedure was repeated at the end of 1st exposure but proceeding from replacing the used mixture with a new solution in the tympanic bulla as described above before the 2nd exposure. Animals with biotin-FITC solution applying to the RWM by soaking it in the tympanic bulla for the same period as USM and USM-x2, respectively but without ultrasound irradiation will be defined as round window soaking groups (RWS and RWS-x2, respectively) and served as controls. See also page 441 which states use of 3 W/cm2 US intensity. See also Figure 1, in which MBs-ultrasound exposure showed in USM-x2 group revealed far more higher FITC intensity compared to that in USM group that received only one exposure. It would have been obvious to one of ordinary skill in the art at the time of the invention to modify the ultrasound exposure such as to include an additional ultrasound exposure as a matter of routine optimization with a reasonable expectation of success in providing drug delivery having favorable conditions as it was shown that increased MR ultrasound exposure resulted in increased delivery.
Claim(s) 13 is rejected under 35 U.S.C. 103 as being unpatentable over Liao et al. (US 2015/0056273, hereinafter Liao I), in view of Liao et al. (2012 IEEE International Ultrasonics Symposium Proceedings, p. 440-443, hereinafter Liao II), in further view of Voorhies et al. (US 2005/0060012).
Liao I teaches an external type microbubble ultrasound contrast agent may employ a medium, either aqueous or a gel form, and contain microbubbles of a specific particle size and at a specific concentration. The material of the microbubbles may be albumin, liposomes, polymers, copolymers or mixtures of the aforementioned material or a combination of those above. The material of the microbubbles may also include pentose and/or hexose. A method for preparing an external type microbubbles ultrasound contrast agent of topical uses is also provided. The microbubble ultrasound contrast agent may have microbubbles with different particle sizes by adjusting the percentage of the medium and the material of the microbubbles the mixed solution and followed by the same ultrasonic oscillating steps which oscillating the mixed solution for about 100 to about 140 seconds. The external type microbubble ultrasound contrast agent may be applied in conjunction with the application of mechanical oscillation waves. Through a series of swelling and shrinking processes induced by the oscillation energy of the mechanical oscillation waves, the microbubbles burst or destructed and the generated energy and shock waves lead to minor damages of cells or tissues, which further strengthen the absorption of applied chemicals or small molecules, also the microbubbles with different particle sizes may lead to different penetration depth for the applied chemicals or small molecules. The commonly used energy source of the mechanical oscillation waves may be a source of an optical energy or acoustic energy, such as an ultrasound source or a laser source. The external type microbubble ultrasound contrast agent of the present invention, suitable for applying onto a local region of the body surface of the living body, may be used in combination with the mechanical wave(s) generated by the mechanical oscillating energy source to cause the microbubbles in the external type microbubble ultrasound contrast agent bursting to produce energy and shock waves. The energy and the shock waves from microbubble bursting cause minor and reversible damages on the contact area of the skin surface or mucous membrane, thereby increasing the percutaneous absorption of chemicals or small molecules. The microbubble ultrasound contrast agent may be widely used in medical or beauty fields, to help strengthen the absorption of painkillers after surgery or the absorption of beauty care ingredients (paragraph 0006).
The microbubble contrast agent can enhance the delivery of the chemical and promote the absorption of the drug or small molecules (paragraph 0103).
The preparation of the microbubble contrast agent is set forth in paragraphs 0062+, including providing an isotonic saline solution comprising microbubbles, (Component A, the microbubble liquid); a drug or other chemical to be delivered (Component B) and Component B is used a diluent to dilute Component A 2-1000 times and the composition obtained after dilution is applied to the surface of the body.
The concentration of the particles is preferably 2x106 – 2x108 particles/ml (paragraph 0066).
The size of the microbubbles is 0.5 to 3.7 micrometers (paragraph 0059).
The material of the microbubbles is albumin, liposomes, polymers, etc. (paragraph 0059).
The external use microbubble contrast agent of the present invention may be used in the ear treatments. The microbubble contrast agent of this invention is mixed with the dye and/or one or more medical ingredients and administrated to the inner ear of guinea pigs. The administration of the mixtures may be conducted in different ways to test the delivery efficiency of the dye or the ingredient (paragraph 0099).
The animals used in the test are 60 guinea pigs with the normal Preyer's reflex to the sound(s) and are divided into three groups with the following experimental conditions: (1) the tympanic bullae of 24 guinea pigs are filled with the microbubble ultrasound contrast agent mixed the dye indicator and applied with the ultrasound (paragraph 0101).
Sonoporation Gene Transfection System (ST2000V, NepaGene, Japan), with a probe size of 6 mm and the waveform of square waves. In the experiments, the ultrasound is operated at a frequency of 1 MHz, a duty cycle of 50%, energy of 3 W/cm.sup.2, is applied for 1 minute. In the experiments, the probe is placed on the body surface facing the round window membrane with a distance of 5 mm (paragraph 0102).
FIG. 12 shows the results of the delivery efficiency using different administration approaches of the microbubble contrast agent in the inner ear treatment experiments.
Compared to the control group of delivering the dye or drug into the inner ear through the diffusion effect, the experimental results indicate that the ultrasound used together with the microbubble ultrasound contrast agent can enhance the drug delivery efficiency. In addition, in order to deliver gentamycin into the inner ear, the microbubbles ultrasound contrast agent of this invention is used along with the application of the ultrasound. By using such approach, the concentration of gentamycin delivered into the cochlear tissues is significantly higher than that of the control group without applying the ultrasound. Hence it is confirmed that the microbubble contrast agent can enhance the delivery of the chemical and promote the absorption of the drug or small molecules.
FIGS. 13A-13F show the delivery results of the green dye indicator entering into the round window membrane cells of the inner ear under different administration approaches. FIGS. 13A-136C show the delivery results of the experimental groups using the ultrasound microbubble contrast agent mixed with the green dye indicator and operated with the ultrasound. FIGS. 13D-13F show the delivery results of the control groups using the ultrasound microbubble contrast agent mixed with the green dye indicator but without applying the ultrasound (through the diffusion effect). Compared the results of little or no green dye entering into the round window membrane cells in FIGS. 13D-136F, the results of FIGS. 13A-13C show much more green dyes entering into the round window membrane cells (paragraph 0103).
FIGS. 13A-13F show the delivery results of the green dye indicator entering into the round window membrane cells of the inner ear under different administration approaches. FIGS. 13A-136C show the delivery results of the experimental groups using the ultrasound microbubble contrast agent mixed with the green dye indicator and operated with the ultrasound (paragraph 0104).
Liao II teaches that direct drug delivery into inner ear can be achieved by three approaches: (1) diffusion of drug through round window; (2) injection of drug through round window; (3) cochleostomy or canalostomy. The latter two approaches are invasive and have the risk of hearing loss and vertigo. Drug diffusion through round window is only the noninvasive approach. However, how to enhance drug diffusion and how to noninvasively promote drug delivery through round window are two issues that need to be investigated. In this study, we target on the practical application of microbubbles (MBs)-ultrasound on increasing the wound window membrane (RWM) permeability for facilitating drug or medication delivering into the inner ear. Using biotin-FITC conjugates (biotin-FITC) as delivered agents and performed on guinea pigs animal models, we showed that the MBs-ultrasound exposure can greatly improve the inner ear system utility of the biotin-FITC delivery via RWM at different kinds of approaches about 3.5 to 38 times compare to that solely soaking biotin-FITC around the RWM for spontaneous diffusion. In addition, significant enhancement of hair cells uptake of gentamicin was demonstrated in animals whose tympanic bullas were soaked with MBs-mixed gentamicin−Texas Red or gentamicin and exposed to ultrasound. Furthermore, the increased permeability of RWM resulted from acoustic cavitation of MBs could also be visualized immediately following ultrasound exposure by using Alexa Fluor 488-conjugated phalloidin as a tracer. Most importantly, such applications were shown without resulting damage to the integrity of RWM or deterioration of the hearing thresholds assessed by auditory brainstem responses, suggesting this MBs-ultrasound not only benefits in developing therapeutic strategies for inner ear diseases, but also help in providing a more precise and well-controlled release for medications passing through the RWM (abstract).
Albumin-shelled MBs were prepared and mixed with gentamicin-Texas red conjugate. A fenestration allows exposing the round window, loading the mixture of MBs, and delivering reagents into the middle ear cavity for sonication.
A fenestration (approximately 4 mm in diameter) was created in the tympanic bulla by drilling with diamond burrs. This fenestration allows exposing the round window, loading the mixture of MBs, and delivering reagents into the middle ear cavity for sonication (page 441).
It was shown that ultrasound-aided microbubbles promote the delivery of biotin to the inner ear. Cavitation of MBs-ultrasound results in increase of round window membrane permeability without causing complete cell damage. Microbubble-ultrasound exposure did not deteriorate hearing acuity. The MBs-ultrasound system appears to be safe in our guinea pigs model. Even though, a comprehensive histology study of the inner ear following MBs-ultrasound exposure is needed before final conclusions concerning safety issue can be made (page 442).
Accordingly, Liao I and II teach a method of delivering drugs to inner ear facilitated by microbubbles comprising providing a microbubble-drug mixture and applying to the middle ear cavity and sonication by ultrasonic waves to produce cavitation so as to increase permeability of the round window membrane allowing the drug in the microbubble-drug mixture to penetrate the round window membrane. However, Liao I recites that microbubbles are in direct contact with the ultrasound probe to induce cavitation under the ultrasound energy (paragraph 0057), rather than in indirect contact as claimed. Liao I and II do not specifically recite wherein mechanical waves are in contact with a conductive glue applied to the skull skin behind the ear corresponding to the tympanic bulla.
Voorhies teaches noninvasive ultrasound, in which a method and system is provided to induce mild hypothermia in a patient through controlled heating of the preoptic anterior hypothalamus (POAH) in conjunction with cooling of patient's body. The system employs an ultrasound transducer that may be positioned extracorporeally to a patient skull for emitting ultrasound energy to the POAH. The ultrasound energy heats the POAH to inhibit thermoregulatory responses of the body such that a cooling means may more effectively cool bodily tissue in order to reduce a patient's core body temperature (abstract).
The frequency ranges, power, intensity and pulse length settings generally associated with physiotherapy ultrasound are utilized with the current PAOH heating system 5. However, it will be appreciated that depending on the settings utilized, diagnostic and/or surgical ultrasound devices may also be utilized. In this regard, it is believed that the full medical ultrasound frequency range and power levels may be utilized with the present system 5. In any case, the settings of the transducer device 20 may be set at a first predetermined level for initial POAH heating and those settings (e.g., power, intensity, pulse length and/or frequency) may be subsequently increased or decreased depending on body temperature feedback, as will be discussed herein. In order to heat the POAH, the transducer device 20 is semi-permanently affixed to the patient's skull using an adhesive and/or headband to maintain proper focus on the POAH (paragraph 0044-45).
In addition to a fluid-containing layer 120, the back pads 100 may each further include a conformable, thermally conductive layer 160 for contacting the skin of a patient. In this regard, the conformable layer 160 may provide an adhesive surface 162 for enhancing the pad-to-skin interface. Preferably, the adhesive surface 162 extends across a major portion (e.g. substantially all) of the pads 100a, 100b. A release liner 170 may also be provided on the adhesive surface 162 for removal prior to use (paragraph 0052).
It would have been obvious to one of ordinary skill in the art at the time of the invention to provide the oscillation wave source in a non-invasive manner in contact with a conductive glue applied to the skull skin behind the ear when the teachings of Liao I and II are taken in view of Voorhies. One would have been motivated to do so, with a reasonable expectation of success, because Liao is concerned with non-invasive promotion of drug delivery through round window, including aversion to interrupting the inner ear structure (Liao II, page 440), and Voorhies teaches non-invasive ultrasound can be applied by affixed to a patient's skull using an adhesive for focus. It would have been further obvious to focus the ultrasound corresponding to the tympanic bulla, as Liao I teaches that the tympanic bullae of guinea pigs are filled with the microbubble composition.
With regard to the amended claim limitation “wherein a 3 W/cm2 ultrasound is applied to the skull skin behind the ear corresponding to the tympanic bulla 3 times and 1 minute each time, and the microbubble-drug mixture in the middle ear cavity is replaced every time to ensure that the microbubble-drug mixture produces the cavitation each time,” see page 442 of Liao II. To determine the efficiency of MBs-ultrasound-mediated delivery in the inner ear system, biotin-FITC conjugates (biotin-FITC) were used as delivered agent. Animals were divided into several experimental groups. In ultrasonic microbubble (USM) group, tympanic bulla was filled with 200 μ1 mixture of biotin-FITC (40 ug/ml) and MBs, while in ultrasonic (US) group only 200 μ1 biotin-FITC solution (40 ug/ml) was filled. Ultrasound exposures were then applied to both groups for 1 min. In USM-x2 and US-x2 groups, the same irradiation procedure was repeated at the end of 1st exposure but proceeding from replacing the used mixture with a new solution in the tympanic bulla as described above before the 2nd exposure. Animals with biotin-FITC solution applying to the RWM by soaking it in the tympanic bulla for the same period as USM and USM-x2, respectively but without ultrasound irradiation will be defined as round window soaking groups (RWS and RWS-x2, respectively) and served as controls. See also page 441 which states use of 3 W/cm2 US intensity. See also Figure 1, in which MBs-ultrasound exposure showed in USM-x2 group revealed far more higher FITC intensity compared to that in USM group that received only one exposure. It would have been obvious to one of ordinary skill in the art at the time of the invention to modify the ultrasound exposure such as to include an additional ultrasound exposure as a matter of routine optimization with a reasonable expectation of success in providing drug delivery having favorable conditions as it was shown that increased MR ultrasound exposure resulted in increased delivery.
Response to arguments
Applicant argues that in terms of the amendments to claim 1, Liao I, II, Noda and Shih fail to teach the claimed feature that “a 3 W/cm2 ultrasound is applied to the external ear canal 3 times and 1 minute each time, and the microbubble-drug mixture in the middle ear cavity is replaced every time to ensure that the microbubble-drug mixture could produce the cavitation each time”. Applicant further argues that Liao I, II, and Voorhies fail to teach the claimed feature and therefore claim 13 as amended herein are non-obvious over the cited references.
Applicant’s arguments have been fully considered but are not found to be persuasive. It is respectfully submitted that with regard to the amended claim limitation “wherein a 3 W/cm2 ultrasound is applied to the skull skin behind the ear corresponding to the tympanic bulla 3 times and 1 minute each time, and the microbubble-drug mixture in the middle ear cavity is replaced every time to ensure that the microbubble-drug mixture produces the cavitation each time,” see page 442 of Liao II. To determine the efficiency of MBs-ultrasound-mediated delivery in the inner ear system, biotin-FITC conjugates (biotin-FITC) were used as delivered agent. Animals were divided into several experimental groups. In ultrasonic microbubble (USM) group, tympanic bulla was filled with 200 μ1 mixture of biotin-FITC (40 ug/ml) and MBs, while in ultrasonic (US) group only 200 μ1 biotin-FITC solution (40 ug/ml) was filled. Ultrasound exposures were then applied to both groups for 1 min. In USM-x2 and US-x2 groups, the same irradiation procedure was repeated at the end of 1st exposure but proceeding from replacing the used mixture with a new solution in the tympanic bulla as described above before the 2nd exposure. Animals with biotin-FITC solution applying to the RWM by soaking it in the tympanic bulla for the same period as USM and USM-x2, respectively but without ultrasound irradiation will be defined as round window soaking groups (RWS and RWS-x2, respectively) and served as controls. See also page 441 which states use of 3 W/cm2 US intensity. See also Figure 1, in which MBs-ultrasound exposure showed in USM-x2 group revealed far more higher FITC intensity compared to that in USM group that received only one exposure. It would have been obvious to one of ordinary skill in the art at the time of the invention to modify the ultrasound exposure such as to include an additional ultrasound exposure as a matter of routine optimization with a reasonable expectation of success in providing drug delivery having favorable conditions as it was shown that increased MR ultrasound exposure resulted in increased delivery.
Conclusion
No claims are allowed at this time.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/LHS/
/Michael G. Hartley/Supervisory Patent Examiner, Art Unit 1618