Organic acid salt of nicotinamide riboside, composition including organic acid salt, and preparation methods of organic acid salt and composition

Abstract

An organic acid salt of nicotinamide riboside (NR) is provided, where an organic acid is selected from the group consisting of malic acid, citric acid, royal jelly acid, and the like, and a molar ratio of the NR to the organic acid is 1:2 or 1:1. A compound composition of an organic acid salt of NR and a carrier is further provided, where the carrier is selected from the group consisting of niacin, glutamic acid, royal jelly acid, nervonic acid, microcrystalline cellulose (MCC), and apple cider vinegar powder. The present disclosure adopts an organic acid with strong acidity, and such an organic acid can form an intimate ion pair with NR and show some hydrophobicity, which improves the stability of NR. The carrier can prevent moisture penetration and improve the water resistance of the organic acid salt.

Claims

1. An organic acid salt of nicotinamide riboside (NR), wherein an organic acid is citric acid or malic acid; a molar ratio of the NR to the organic acid is 1:2.

2. A preparation method of the organic acid salt of NR according to claim 1, comprising: under a protection of nitrogen, dissolving the NR in methanol, adding the organic acid to obtain a first resulting mixture, and stirring the first resulting mixture; adding methyl tert-butyl ether (MTBE) or ethyl acetate to obtain a second resulting mixture, and further stirring the second resulting mixture; and filtering, washing, and drying to obtain a product.

3. A composition of the organic acid salt of NR, comprising the organic acid salt of NR and a carrier, wherein the carrier is selected from the group consisting of malic acid, tannic acid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), caffeic acid, trans-cinnamic acid, trans-4-hydroxy-cinnamic acid, lactic acid, monosodium citrate, disodium citrate, citric acid, chlorogenic acid, gluconic acid, ferulic acid, royal jelly acid, nervonic acid, chicoric acid, rosmarinic acid, carnosic acid, niacin, adipic acid, lauric acid, monopotassium glycyrrhizinate (MPG), folic acid, chondroitin sulfate (CS), potassium hydrogen tartrate, salicylic acid, glycine, glutamic acid, alanine, arginine, leucine, isoleucine, valine, cystine, cysteine, methionine, threonine, serine, phenylalanine, tyrosine, tryptophan, proline, hydroxyproline, and aspartic acid; and the organic acid salt is the organic acid salt according to claim 1.

4. The composition of the organic acid salt of NR according to claim 3, wherein the carrier is selected from the group consisting of niacin, glutamic acid, royal jelly acid, and nervonic acid, and the organic acid salt is a malate of NR.

5. A composition of the organic acid salt of NR, comprising the organic acid salt of NR and a carrier, wherein the carrier is selected from the group consisting of microcrystalline cellulose (MCC) and apple cider vinegar powder; and the organic acid salt is the organic acid salt according to claim 1.

6. A preparation method of the composition according to claim 3, comprising: under a protection of nitrogen, mixing and grinding the organic acid salt of NR and the carrier.

7. The preparation method of the composition according to 6, wherein the carrier is selected from the group consisting of niacin, glutamic acid, royal jelly acid, and nervonic acid, and the organic acid salt is a malate of NR.

8. A preparation method of the composition according to 5, comprising: under a protection of nitrogen, mixing and grinding the organic acid salt of NR and the carrier.

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

(1) The present disclosure is further described below with reference to specific examples. The parts below all refer to parts by weight.

EXAMPLE 1

(2) Preparation of a citrate of NR (1:1)

(3) Under the protection of nitrogen and at a temperature of −10° C. to −5° C., NR (0.0220 mol, 1 eq) was dissolved in 60 mL of methanol, then anhydrous citric acid (0.0264 mol, 1.2 eq) was added, and a resulting mixture was stirred for 2 h; and then 75 mL of anhydrous MTBE was added, and a resulting mixture was further stirred for 30 min, filtered under the protection of nitrogen, rinsed with anhydrous diethyl ether, and dried at a temperature below −5° C. to obtain a product (0.0123 mol). In the product (NRX), a molar ratio of NR to citric acid (X) was 1:1; and a yield was 55.91%.

(4) Characterization data:

(5) HNMR(400 MHz,MeOD):δ9.72 (s,1H), 9.42-9.43 (d,1H), 9.01-9.03 (d,1H), 8.25-8.30 (m,1H), 6.18-6.19 (d,1H), 4.42-4.464 (m,2H), 4.30-4.32 (t,1H), 4.01-4.05 (dd,1H), 3.85-3.89 (dd,1H), 2.67-2.79 (q,4H);

(6) MS(ESI+):254.96[M-1],MS(ESI−): 191.12[M-1];

(7) IR (KBr) v.sub.max 3412, 2940, 2375, 1692, 1612, 1516, 1395, 1232, 1098, 894, 677, 622 cm.sup.−1.

EXAMPLE 2

(8) Preparation of a citrate of NR (1:2)

(9) Under the protection of nitrogen and at a temperature of −10° C. to −5° C., NR (0.0220 mol, 1 eq) was dissolved in 60 mL of methanol, then anhydrous citric acid (0.0495 mol, 2.25 eq) was added, and a resulting mixture was stirred for 2 h; and then 150 mL of anhydrous MTBE was added, and a resulting mixture was further stirred for 30 min, filtered under the protection of nitrogen, rinsed with anhydrous diethyl ether, and dried at a temperature below −5° C. to obtain a product (0.0125 mol). In the product (NRX.sub.2), a molar ratio of NR to citric acid (X) was 1:2; and a yield was 56.82%.

(10) Characterization data:

(11) HNMR(400 MHz,MeOD):δ9.73 (s,1H), 9.43-9.45 (d,1H), 9.04-9.06 (d,1H), 8.27-8.32 (m,1H), 6.20-6.21 (d,1H), 4.43-4.45 (m,2H), 4.30-4.33 (t,1H), 4.02-4.05 (d,1H), 3.85-3.89 (d,1H), 2.74-2.86 (q,8H);

(12) MS(ESI+):254.97[M-1],MS(ESI−): 191.16[M-1];

(13) IR (KBr) v.sub.max 3425, 2929, 2370, 1697, 1624, 1516, 1394, 1219, 1097, 893, 678, 623 cm.sup.−1.

(14) When the NR and the anhydrous citric acid were fed in a molar ratio of 1:3.3, characterization data of an obtained product were the same, indicating that the product was a product (NRX.sub.2) in which a molar ratio of NR to citric acid (X) was 1:2.

EXAMPLE 3

(15) Preparation of a malate of NR (1:1)

(16) Under the protection of nitrogen and at a temperature of −10° C. to −5° C., NR (0.0195 mol, 1 eq) was dissolved in 60 mL of methanol, then anhydrous malic acid (0.0234 mol, 1.2 eq) was added, and a resulting mixture was stirred for 2 h; and then 100 mL of anhydrous MTBE was added, and a resulting mixture was further stirred for 30 min, filtered under the protection of nitrogen, rinsed with anhydrous diethyl ether, and dried at a temperature below −5° C. to obtain a product (0.01334 mol). In the product (NRX), a molar ratio of NR to malic acid (X) was 1:1; and a yield was 57%.

(17) Characterization data:

(18) HNMR(400 MHz,MeOD): δ9.72 (s,1H), 9.42-9.44 (d,1H), 9.02-9.04 (d,1H), 8.25-8.29 (t,1H), 6.18-6.19 (d,1H), 4.41-4.44 (m,2H), 4.26-4.30 (m,2H), 3.99-4.03 (dd,1H), 3.83-3.87 (dd,1H), 2.49-2.80 (dd,2H);

(19) MS(ESI+):254.96[M-1],MS(ESI−):133.04[M-1];

(20) IR (KBr) v.sub.max3379, 2937, 1691, 591, 1100, 677, 6 cm.sup.−.

EXAMPLE 4

(21) Preparation of a malate of NR (1:2)

(22) Under the protection of nitrogen and at a temperature of −10° C. to −5° C., NR (0.0195 mol, 1 eq) was dissolved in 45 mL of methanol, then anhydrous malic acid (0.043875 mol, 2.25 eq) was added, and a resulting mixture was stirred for 2 h; and then 80 mL of anhydrous MTBE was added, and a resulting mixture was further stirred for 30 min, filtered under the protection of nitrogen, rinsed with anhydrous diethyl ether, and dried at a temperature below −5° C. to obtain a product (0.0118 mol). In the product (NRX.sub.2), a molar ratio of NR to malic acid (X) was 1:2; and a yield was 60.51%.

(23) Characterization data:

(24) HNMR(400 MHz,MeOD): δ9.73(s,1H), 9.43-9.45 (d,1H), 9.04-9.06 (d,1H), 8.27-8.31 (m,1H), 6.19-6.20 (d,1H), 4.43-4.45 (m,2H), 4.31-4.34 (m,3H), 4.01-4.05 (dd,1H), 3.85-3.89 (dd,1H), 2.53-2.83 (dd,4H);

(25) MS(ESI+):254.94[M-1],MS(ESI−):133.03[M-1];

(26) IR (KBr) v.sub.max3409, 2940, 1698, 1580, 1411, 1293, 1181, 1098, 1028, 658 cm.sup.−1.

(27) When the NR and the anhydrous malic acid were fed in a molar ratio of 1:3.3, characterization data of an obtained product were the same, indicating that the product was a product (NRX.sub.2) in which a molar ratio of NR to malic acid (X) was 1:2.

EXAMPLE 5

(28) Preparation of a royal jelly acid salt of NR (1:2)

(29) Under the protection of nitrogen and at a temperature of −10° C. to −5° C., NR (0.0400 mol, 1 eq) was dissolved in 60 mL of methanol, then royal jelly acid (0.0860 mol, 2.15 eq) was added, and a resulting mixture was stirred for 2 h; and then 120 mL of anhydrous ethyl acetate was slowly added, and a resulting mixture was further stirred for 30 min, filtered under the protection of nitrogen, rinsed with anhydrous diethyl ether, and dried at a temperature below −5° C. to obtain 12.5 g of a product (0.02 mol). In the product (NRX.sub.2), a molar ratio of NR to royal jelly acid (X) was 1:2; and a yield was 50%.

(30) Characterization data:

(31) HNMR(400 MHz,MeOD): δ9.73 (s,1H), 9.44 (s,1H), 9.04-9.05 (d,1H), 8.30 (m,1H), 6.58-6.66 (m,2H), 6.18 (m,1H), 5.81-5.85 (d,2H), 4.92 (m,2H), 4.31-4.43 (t,1H), 3.85-4.05 (dd,2H), 3.54-3.57 (t,4H), 2.13-2.18 (m,4H), 1.51-1.56 (m,4H), 1.45-1.49 (m,4H), 1.37 (m,12H);

(32) MS(ESI+):254.96[M-],MS(ESI−): 185.23[M-1];

(33) IR (KBr) v.sub.max3384, 2924, 1705, 1654, 1555, 1421, 1389, 1187, 1098, 1053, 977, 869, 677 cm.sup.−1.

EXAMPLE 6

(34) Preparation of a malate of NR with niacin as a carrier

(35) Under the protection of nitrogen, 0.01 mol of the malate of NR in Example 4 was added to 0.01 mol of niacin, and a resulting mixture was ground at a low temperature (about 16° C., the same below) for about 10 min until the mixture had a particle size of about 200 mesh to obtain 6.4 g of a mixture of malate and nicotinate of NR.

(36) IR (KBr) v.sub.max3409, 2927, 1695, 1584, 1398, 1319, 1101, 1028, 747, 677, 636 cm.sup.−1.

EXAMPLE 7

(37) Preparation of a malate of NR with glutamic acid as a carrier

(38) Under the protection of nitrogen, 0.01 mol of the malate of NR in Example 4 was added to 0.01 mol of glutamic acid, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 6.65 g of a mixture of malate and glutamate of NR.

(39) IR (KBr) v.sub.max3415, 2937, 1692, 1593, 1404, 1092, 1028, 670 cm.sup.−1.

EXAMPLE 8

(40) Preparation of a malate of NR with royal jelly acid as a carrier

(41) Under the protection of nitrogen, 0.01 mol of the malate of NR in Example 4 was added to 0.01 mol of royal jelly acid, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 7.05 g of a mixture of malate and royal jelly acid salt of NR.

(42) IR (KBr) v.sub.max3437, 2934, 1698, 1651, 1401, 1095, 684 cm.sup.−1.

EXAMPLE 9

(43) Preparation of a malate of NR with nervonic acid as a carrier

(44) Under the protection of nitrogen, 0.01 mol of the malate of NR in Example 4 was added to 0.01 mol of nervonic acid, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 8.85 g of a mixture of malate and nervonic acid salt of NR.

(45) IR (KBr) v.sub.max3415, 2924, 1695, 1465, 1418, 1290, 1098, 728, 674 cm.sup.−1.

EXAMPLE 10

(46) Preparation of a malate of NR with MCC as a carrier

(47) Under the protection of nitrogen, 1 g of the malate of NR in Example 4 was added to 1 g of MCC, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 2 g of a mixture of malate of NR and MCC.

(48) IR (KBr) v.sub.max3425, 2930, 1688, 1644, 1513, 1395, 1095, 1018, 667 cm.sup.−1.

EXAMPLE 11

(49) Preparation of a malate of NR with MCC as a carrier

(50) Under the protection of nitrogen, 1 g of the malate of NR in Example 3 was added to 1 g of MCC, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 2 g of a mixture of malate of NR and MCC.

(51) IR (KBr) v.sub.max3365, 2906, 1695, 1591, 1396, 1099, 675 cm.sup.−1.

EXAMPLE 12

(52) Preparation of a citrate of NR with MCC as a carrier

(53) Under the protection of nitrogen, 1 g of the citrate of NR in Example 2 was added to 1 g of MCC, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 2 g of a mixture of citrate of NR and MCC.

(54) R (KBr) v.sub.max3437, 2927, 2378, 1740, 1698, 1641, 1513, 1398, 1111, 897, 671 cm.sup.−1.

EXAMPLE 13

(55) Preparation of a citrate of NR with MCC as a carrier

(56) Under the protection of nitrogen, 1 g of the citrate of NR in Example 1 was added to 1 g of MCC, and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 2 g of a mixture of citrate of NR and MCC.

(57) IR (KBr) v.sub.max 3415, 2921, 2368, 1698, 1625, 1513, 1401, 1098, 894, 670, 619 cm.sup.−1.

EXAMPLE 14

(58) Preparation of a malate of NR with apple cider vinegar powder as a carrier

(59) Under the protection of nitrogen, 1 g of the malate of NR in Example 4 was added to 1 g of apple cider vinegar powder (10%), and a resulting mixture was ground at a low temperature for about 10 min until the mixture had a particle size of about 200 mesh to obtain 2 g of a mixture of malate of NR and apple cider vinegar powder.

(60) IR (KBr) v.sub.max3406, 2930, 1692, 1584, 1408, 1092, 1028, 674 cm.sup.−1.

COMPARATIVE EXAMPLE

(61) Preparation of a chlorine salt of NR

(62) Under the protection of nitrogen and at a temperature of −10° C. to −5° C., NR (0.0220 mol, 1 eq) was dissolved in 50 mL of methanol, then 10 g of a 17% hydrogen chloride-methanol solution was added dropwise, and a resulting mixture was stirred for dissolution; then 1 g of activated carbon was added, and a resulting mixture was further stirred for 1 h and filtered; a resulting filtrate was added to 150 ml of anhydrous MTBE, and a resulting mixture was further stirred for 30 min, filtered under the protection of nitrogen, rinsed with anhydrous diethyl ether, and dried at a temperature below −5° C. to obtain 8 g of a product.

(63) The method in Example 10 was used to prepare a chlorine salt of NR with MCC as a carrier.

(64) Characterization data:

(65) HNMR(400 MHz,MeOD): 9.72 (s,1H), 9.44-9.46 (d,1H), 9.05-9.07 (d,1H), 8.29-8.33 (t,1H), 6.22-6.23 (d,1H), 4.45-4.47 (t,1H), 4.41-4.43 (q,1H), 4.31-4.33 (t,1H), 3.99-4.03 (d,1H), 3.84-3.88 (d,1H).

(66) IR (KBr) v.sub.max3336, 2935, 1687, 1616, 1400, 1100, 675 cm.sup.−1.

(67) Stability test

(68) I. Preparation of samples

(69) 1. The products of Examples 1 to 4, Examples 10 to 13, and the comparative example were each dispensed into 13 bottles, each of 50 mg; and the bottles were filled with nitrogen and sealed. One of the bottles (initial sample) was first tested, and the remaining 12 bottles were divided into two groups (each with 6 bottles) and stored. The bottles in each group were numbered 1 to 6 according to a test sequence. The two groups of bottles were stored at 2° C. to 8° C. and −20° C., respectively; and one sample was tested every month.

(70) 2. The products of Examples 6 to 9 and Example 14 were each dispensed into 7 bottles, each of 50 mg; and the bottles were filled with nitrogen and sealed. One of the bottles (initial sample) was first tested, and the remaining 6 bottles were numbered 1 to 6 according to a test sequence and then stored at 2° C. to 8° C. One sample was tested every month.

(71) II. Preparation of test sample solutions

(72) One bottle of sample was taken at a time and prepared into a 5 mL or 10 mL solution using a 5 mL or 10 mL volumetric flask, and then the solution was filtered through a membrane to be ready for purity test by HPLC.

(73) III. HPLC test

(74) Mobile phase: isocratic elution: 5% water (0.1% formic acid)+95% methanol (0.1% formic acid)

(75) Wavelength: 254 nm

(76) Temperature and humidity: 23.0° C. and 54% RH

(77) Sample dissolution: dissolution by methanol

(78) Chromatographic column: ODS-2, 4.6 * 250 mm, 5 μm, and constant pressure: 12 Mpa to 13 Mpa

(79) Flow rate: 1.0 mL/min

(80) Injection volume: 5 μL

(81) Running time: ≥15 min IV.

(82) $\mathrm{Decomposition}\mathrm{rate}=\left(\mathrm{purity}\mathrm{of}\mathrm{initial}\mathrm{sample}-\mathrm{purity}\mathrm{of}\mathrm{sample}6\right)/\mathrm{purity}\mathrm{of}\mathrm{initial}\mathrm{sample}\times 100\%$

(83) TABLE-US-00001 TABLE 1 (−20° C.) Exam- Exam- Exam- Exam- Comparative Time (month) ple 1 ple 2 ple 3 ple 4 Example 0 98.930 98.471 98.573 98.818 98.525 1 98.828 98.317 98.691 98.547 98.284 2 98.450 98.079 98.310 98.629 98.087 3 98.190 98.168 98.288 98.316 97.701 4 98.252 97.933 98.076 98.207 97.449 5 97.978 97.806 97.912 98.081 97.575 6 97.814 97.788 97.645 98.100 97.210 Decomposition rate 1.13% 0.69% 0.94% 0.73% 1.33%

(84) TABLE-US-00002 TABLE 2 (2° C. to 8° C.) Exam- Exam- Exam- Exam- Comparative Time (month) ple 1 ple 2 ple 3 ple 4 Example 0 98.930 98.471 98.573 98.818 98.525 1 98.198 98.492 98.482 98.424 98.171 2 98.409 98.240 98.185 98.360 98.206 3 98.171 98.097 98.270 98.298 97.540 4 97.957 97.626 97.946 98.239 97.612 5 97.823 97.704 97.592 97.707 97.384 6 97.670 97.498 97.337 97.816 97.109 Decomposition rate 1.27% 0.99% 1.25% 1.01% 1.44%

(85) It can be seen from Table 1 and Table 2 that the lower the temperature, the better the stability of the salt of NR; and the malate and citrate of NR (in a molar ratio of either 1:1 or 1:2) are both better than the chlorine salt of NR (comparative example), this is because malic acid or citric acid can form an intimate ion pair with NR, which leads to some hydrophobicity and thus improves the stability.

(86) From the comparison of Example 1 with Example 2 and Example 3 with Example 4, it is found that the molar ratio (NR to organic acid) of 1:2 leads to higher stability than the molar ratio (NR to organic acid) of 1:1. In a molar ratio of 1:1, NR and an organic acid are preferably present in the form of an ion pair according to the electron effect. When NR and an organic acid are fed in a molar ratio of 1:2, according to HNMR, it is found that one molecule of the NR and two molecules of the organic acid can exist stably. One molecule of the organic acid forms an ion pair with the NR, and then the other molecule of the organic acid forms a hydrogen bond with basic amide in the NR due to acid-base interaction, thereby achieving acid-base pairing, which helps to improve the stability of the salt. In addition, the two molecules of acid can provide a strongly acidic environment, and the stronger the acidity, the higher the stability of NR. For example, the acidity of NMN is stronger than that of NR, and thus the stability of NMN is much higher than that of NR. Furthermore, when NR and an organic acid are present in a molar ratio of 1:2, an NR content decreases, the influence of free hydroxyl and amide functional groups among molecules is weakened, and the stability increases.

(87) TABLE-US-00003 TABLE 3 (2° C. to 8° C.) Exam- Exam- Exam- Exam- Exam- Time (month) ple 6 ple 7 ple 8 ple 9 ple 14 0 58.585 97.487 91.719 97.812 97.086 1 58.918 96.170 90.593 96.397 95.776 2 58.755 96.818 92.279 97.108 96.931 3 59.925 96.836 91.762 96.870 96.510 4 57.507 96.504 90.745 96.607 96.208 5 58.363 96.701 90.329 95.914 96.350 6 57.150 95.257 90.140 95.128 95.601 Decomposition rate 2.45% 2.29% 1.72% 2.74% 1.53%

(88) After an organic acid salt of NR is compounded with another organic acid or apple cider vinegar powder, the stability is reduced, but still acceptable. Therefore, the addition of an organic acid or apple cider vinegar powder as a carrier does not significantly affect the stability of the organic acid salt of NR. After an organic acid or apple cider vinegar powder is compounded with NR to form a composite nutritional additive, the components can be complementary and coordinated to produce a superimposed effect, which has potential application value.

(89) TABLE-US-00004 TABLE 4 (−20° C.) Exam- Exam- Exam- Exam- Comparative Time (month) ple 10 ple 11 ple 12 ple 13 Example 0 98.759 98.550 98.316 98.694 97.917 1 98.632 98.180 98.440 98.633 97.874 2 98.714 98.442 98.259 98.441 97.656 3 98.557 98.395 98.213 98.527 97.326 4 98.324 98.308 98.007 98.372 97.470 5 98.491 98.273 98.110 98.393 97.319 6 98.425 98.112 98.041 98.238 97.277 Decomposition rate 0.34% 0.45% 0.28% 0.46% 0.65%

(90) TABLE-US-00005 TABLE 5 (2° C. to 8° C.) Exam- Exam- Exam- Exam- Comparative Time (month) ple 10 ple 11 ple 12 ple 13 Example 0 98.759 98.550 98.316 98.694 97.917 1 98.691 98.339 98.282 98.625 97.776 2 98.506 98.031 98.326 98.458 98.015 3 98.239 98.274 98.028 98.083 97.693 4 98.317 97.932 97.805 98.121 97.542 5 98.342 97.786 97.734 97.833 97.380 6 98.251 97.875 97.782 98.017 97.154 Decomposition rate 0.51% 0.68% 0.54% 0.69% 0.78%

(91) After an organic acid salt of NR is compounded with MCC, the organic acid salt of NR with MCC as a carrier exhibits higher stability (comparison of Examples 10 to 13 with Examples 1 to 4). This is because MCC has hydrophobicity and heat resistance, which can prevent moisture penetration. When MCC is added, the molar ratio (NR to organic acid) of 1:2 (Examples 10 and 12) leads to higher stability than the molar ratio (NR to organic acid) of 1:1 (Examples 11 and 13). The malate or citrate of NR compounded with MCC exhibits higher stability than the corresponding chloride salt of NR compounded with MCC (comparative example), which further supports the previous conclusion.

(92) The above described are merely specific implementations of the present disclosure, and the protection scope of the present disclosure is not limited thereto. Any modification or replacement easily conceived by those skilled in the art within the technical scope of the present disclosure should fall within the protection scope of the present disclosure. Therefore, the protection scope of the present disclosure should be subject to the protection scope of the claims.